Hepatitis c virus immunogenic compositions and methods of use thereof

Abstract

The present disclosure provides nucleic acids comprising nucleotide sequences encoding hepatitis C virus (HCV) E1 and / or E2 polypeptides and / or one or more T-cell epitope polypeptides. The nucleic acids can comprise one or more modifications. The nucleic acids can be self-amplifying. The present disclosure provides compositions comprising the nucleic acids. The present disclosure provides methods of inducing an immune response to HCV in an individual, comprising administering to the individual a composition of the present disclosure.

Description

HEPATITIS C VIRUS IMMUNOGENIC COMPOSITIONS AND METHODS OF USE THEREOF CROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 210,722, filed June 15, 2021, U.S. Provisional Patent Application No.63 / 212,877, filed June 21, 2021, U.S. Provisional Patent Application No.63 / 323,434, filed March 24, 2022, and U.S. Provisional Patent Application No. 63 / 345,799, filed May 25, 2022, which applications are incorporated herein by reference in their entirety. INCORPORATION BY REFERENCE OF SEQUENCE LISTING PROVIDED AS A TEXT FILE

[0002] A Sequence Listing is provided herewith as a text file, “UALB- 56WO_Sequence_Listing_ST25.txt” created on June 12, 2022, and having a size of 740 KB. The contents of the text file are incorporated by reference herein in their entirety. INTRODUCTION

[0003] Hepatitis C virus (HCV) is a blood-borne pathogen that is estimated to infect 150-200 million people worldwide. Infection by HCV may be non-symptomatic, and can be cleared by patients, sometimes without medical intervention. However, the majority of patients develop a chronic HCV infection, which may lead to liver inflammation, scarring, and even to liver failure or liver cancer. In the United States alone, over 3 million people have a chronic infection.

[0004] The HCV virion contains a positive-sense single stranded RNA genome of about 9.5 kb. The genome encodes a single polyprotein of 3,010 to 3,030 amino acids. The structural proteins comprise a core protein forming the viral nucleocapsid and two envelope glycoproteins, E1 and E2.

[0005] There is a need in the art for compositions and methods for inducing immune responses to HCV. SUMMARY

[0006] The present disclosure provides nucleic acids comprising nucleotide sequences encoding hepatitis C virus (HCV) E1 and / or E2 polypeptides and / or one or more T-cell epitope polypeptides. The nucleic acids can comprise one or more modifications. The nucleic acids can be self-amplifying. The present disclosure provides compositions comprising the nucleic acids. The present disclosure provides methods of inducing an immune response to HCV in an individual, comprising administering to the individual a composition of the present disclosure.BRIEF DESCRIPTION OF THE DRAWINGS

[0007] FIG.1A-1C provide an amino acid sequence alignment of examples of the core-E1-E2 coding regions of an HCV genotype 1 virus, specifically representative HCV 1A, 1B and 1C genotypes. Genbank database sequences for the coding region core-E1-E2 were aligned using Geneious software v5.6.4. Numbering of amino acids is according to strain NP_671941 (H77). Consensus: SEQ ID NO:1; AVI1a129: SEQ ID NO:2; NP_671491 (H77): SEQ ID NO:3; EU155269: SEQ ID NO:4; EU781810: SEQ ID NO:5; EU781771: SEQ ID NO:6; AB250610: SEQ ID NO:7; EU781752: SEQ ID NO:8; EU781759: SEQ ID NO:9; EF407439: SEQ ID NO:10; EF407427: SEQ ID NO:11; EU362905: SEQ ID NO:11; EF407413: SEQ ID NO:11; EU781808: SEQ ID NO:12; EU78170: SEQ ID NO:13; AJ238799 (Con1): SEQ ID NO:14; AAK97744: SEQ ID NO:15; AF139594: SEQ ID NO:16; AF176573: SEQ ID NO:17; BAA19625: SEQ ID NO:18; BAA25076: SEQ ID NO:19; BAC54896: SEQ ID NO:20; BAD91386: SEQ ID NO:21; BAF46764: SEQ ID NO:22; BAG30950: SEQ ID NO:22; CAB41951: SEQ ID NO:23; AAK95832: SEQ ID NO:24; AAT69968: SEQ ID NO:25; and BAA03581: SEQ ID NO:26.

[0008] FIG.2A-2C provide an alignment of amino acid sequences of the core-E1-E2 coding region of representative HCV 2A and HCV2B subtypes. Genbank database sequences for the coding region core- E1-E2 were aligned using Geneious software v5.6.4. The amino acid numbering depicted is in accordance with the common HCV strains: D00994 (HC-J6) for HCV 2A; ABO47639 (JFH1) and HPCJ8G-J8 (J8) for HCV2A and HCV2B, respectively. Consensus: SEQ ID NO: 27; D00994 (HC-J6): SEQ ID NO: 28; AB047639 (JFH1): SEQ ID NO:29; AB047645: SEQ ID NO:30; AF238482: SEQ ID NO:31; AY746460: SEQ ID NO:32; HPCPOLP: SEQ ID NO:28; NC_009823: SEQ ID NO:34; HPCJ8G HC-J8: SEQ ID NO:35; AY232730: SEQ ID NO:36; AY232747: SEQ ID NO:37; AB030907: SEQ ID NO38; and DQ430817: SEQ ID NO:39.

[0009] FIG.3A-3C provide an amino acid sequence alignment of the core-E1-E2 coding region for representative HCV 3A, 3B and 3K genotypes. Genbank database sequences for the coding region core- E1-E2 were aligned using Geneious software v5.6.4. Consensus: SEQ ID NO:40; AVI3a177: SEQ ID NO:41; ADF97232(S52): SEQ ID NO:42; YP_0014696: SEQ ID NO:43; CAA54244: SEQ ID NO:44; AAC03058: SEQ ID NO:45; AAY29642: SEQ ID NO:46; ABD85062: SEQ ID NO:47; ABD85063: SEQ ID NO:47; ABD97104: SEQ ID NO:48; BAA06044: SEQ ID NO:49; BAA08372: SEQ ID NO:50; and BAA09890: SEQ ID NO:51.

[0010] FIG.4 provides amino acid sequences of examples of T-cell epitope polypeptides (“TPs”) that can be encoded by an RNA molecule of the present disclosure (SEQ ID Nos:52-61, respectively).

[0011] FIG.5A-5E provide schematic depictions of various embodiments of a nucleic acid(s) of the present disclosure.

[0012] FIG.6A-6H provide: i) nucleotide sequences encoding exemplary nucleic acids depicted schematically in FIG.5A (FIG.6A-6D) (SEQ ID Nos:62 and 64-66, respectively); and ii) mRNA (FIG. 6E-6H) encoded by the nucleotide sequences provided in FIG.6A-6D (SEQ ID Nos:71-74, respectively).

[0013] FIG.7A-7D provide nucleotide sequences of exemplary nucleic acids depicted schematically in FIG.5B (SEQ ID Nos:79 and 64-66, respectively).

[0014] FIG.8A-8F provide schematic depictions of various embodiments of a nucleic acid(s) of the present disclosure.

[0015] FIG.9A-9C provide nucleotide sequences of exemplary nucleic acids depicted schematically in FIG.8A (SEQ ID Nos:80-82, respectively).

[0016] FIG.10A-10B provide nucleotide sequences of exemplary nucleic acids depicted schematically in FIG.8B (SEQ ID Nos:83-84, respectively).

[0017] FIG.11A-11D provide nucleotide sequences of exemplary nucleic acids depicted schematically in FIG.8C (SEQ ID Nos:85-88, respectively).

[0018] FIG.12A-12C provide nucleotide sequences of exemplary nucleic acids depicted schematically in FIG.8D (SEQ ID Nos:89-91, respectively).

[0019] FIG.13 provides a nucleotide sequence of the exemplary nucleic acid depicted schematically in FIG.8E (SEQ ID No:92).

[0020] FIG.14 provides a nucleotide sequence of the exemplary nucleic acid depicted schematically in FIG.8F (SEQ ID No:93).

[0021] FIG.15A-15G provide examples of suitable internal ribosome entry sites (IRES) (SEQ ID Nos:94-100).

[0022] FIG.16A-16B provide an amino acid sequence (FIG.16A) of TP50-C and a nucleotide sequence (FIG.16B) encoding TP50-C, optimized for expression in human cells (SEQ ID Nos:101 and 67, respectively).

[0023] FIG.17A-17B provide an amino acid sequence (FIG.17A) of TP35-NS3 and a nucleotide sequence (FIG.17B) encoding TP35-NS3, optimized for expression in human cells (SEQ ID Nos:102 and 68, respectively).

[0024] FIG.18A-18B provide an amino acid sequence (FIG.18A) of TP27 and a nucleotide sequence (FIG.18B) encoding TP27, optimized for expression in human cells (SEQ ID Nos:103 and 69, respectively).

[0025] FIG.19A-19B provide an amino acid sequence (FIG.19A) of TP48 and a nucleotide sequence (FIG.19B) encoding TP48, optimized for expression in human cells (SEQ ID Nos:104 and 70, respectively).

[0026] FIG.20A-20B provide an amino acid sequence of the core-E1-E2 coding region for HCV genotype 7a. Amino acid sequence for the coding region core-E1-E2 of genotype 7a (isolate QC69; Genbank: ABN05226.1; SEQ ID NO:105) is shown according to the numbering scheme of the reference strain, NP_671941 (H77).

[0027] FIG.21A-21D provide nucleotide sequences, codon optimized for expression in human cells, encoding HCV core-E1-E2 of genotype 1a (FIG.21A) (SEQ ID NO:106), genotype 1b (FIG.21B) (SEQ ID NO:107), genotype 2a (FIG.21C) (SEQ ID NO:108), and genotype 3a (FIG.21D) (SEQ ID NO:109).

[0028] FIG.22A-22H provide schematic depictions of various embodiments of nucleic acids of the present disclosure.

[0029] FIG.23A-23C provide nucleotide sequences of nucleic acids depicted schematically in FIG. 22A (SEQ ID Nos:110-112, respectively).

[0030] FIG.24A-24I provide nucleotide sequences of nucleic acids depicted schematically in FIG.22A (SEQ ID Nos:113-121, respectively).

[0031] FIG.25A-25L provide nucleotide sequences of nucleic acids depicted schematically in FIG. 22B (SEQ ID Nos:122-133, respectively).

[0032] FIG.26A-26B provide a nucleotide sequence (FIG.26A) encoding an HCV Core-E1-E2 polyprotein of genotype 4 (SEQ ID NO:134); and an amino acid sequence (FIG.26B) of the encoded HCV Core-E1-E2 polyprotein (SEQ ID NO:135).

[0033] FIG.27A-27B provide a nucleotide sequence (FIG.27A) encoding an HCV Core-E1-E2 polyprotein of genotype 5 (SEQ ID NO:136); and an amino acid sequence (FIG.27B) of the encoded HCV Core-E1-E2 polyprotein (SEQ ID NO:137).

[0034] FIG.28A-28B provide a nucleotide sequence (FIG.28A) encoding an HCV Core-E1-E2 polyprotein of genotype 6 (SEQ ID NO:138); and an amino acid sequence (FIG.28B) of the encoded HCV Core-E1-E2 polyprotein (SEQ ID NO:139).

[0035] FIG.29 provides the amino acid sequence of a TP fusion polypeptide (SEQ ID NO:140).

[0036] FIG.30 provides a schematic depiction of an embodiments of nucleic acids of the present disclosure.

[0037] FIG.31A-31C provide examples of nucleotide sequences encoding RNAs, as depicted in FIG. 30 (SEQ ID NOs:141, 144 and 146, respectively).

[0038] FIG.32A-32C provide amino acid sequences of E1 / E2, TP465, and TP156 polypeptides encoded by the nucleotide sequence set forth in FIG.31A (SEQ ID NOs:148-150, respectively).

[0039] FIG.33A-33C provide amino acid sequences of E1 / E2, TP465, and TP48 polypeptides encoded by the nucleotide sequence set forth in FIG.31B (SEQ ID NOs:151-153, respectively).

[0040] FIG.34A-34C provide amino acid sequences of E1 / E2, TP465, and TP65 polypeptides encoded by the nucleotide sequence set forth in FIG.31C (SEQ ID NOs:154-156, respectively). DEFINITIONS

[0041] The term “hepatitis C virus” (“HCV”), as used herein, refers to any one of a number of different genotypes and isolates of hepatitis C virus. Thus, “HCV” encompasses any of a number of genotypes, subtypes, or quasispecies, of HCV, including, e.g., genotype 1, 2, 3, 4, 5, 6, 7, etc. and subtypes (e.g., 1a, 1b, 2a, 2b, 3a, 4a, 4c, etc.), and quasispecies. Representative HCV genotypes and isolates include: HCV- 1, H77, J6, Con1, isolate 1, BK, EC1, EC10, HC-J2, HC-J5; HC-J6, HC-J7, HC-J8, HC-JT, HCT18, HCT27, HCV-476, HCV-KF, “Hunan”, “Japanese”, “Taiwan”, TH, type 1, type 1a, H77 type 1b, type 1c, type 1d, type 1e, type 1f, type 10, type 2, type 2a, type 2b, type 2c, type 2d, type 2f, type 3, type 3a, type 3b, type 3g, type 4, type 4a, type 4c, type 4d, type 4f, type 4h, type 4k, type 5, type 5a, type 6, type 6a, and type 7a.

[0042] The terms "individual," "host," "subject," and "patient" are used interchangeably herein, and refer to a mammal, including, but not limited to, non-human primates (e.g., simians), equines (e.g., horses), rodents (e.g., rats; mice), and humans.

[0043] As used herein, the term “isolated,” in reference to a nucleic acid, refers to a nucleic acid that is in an environment different from that in which the nucleic acid naturally occurs or in which the nucleic acid was synthesized. An isolated nucleic acid can be purified. By “purified” is meant a compound of interest (e.g., a nucleic acid) has been separated from components that accompany it in nature or that are found together with the nucleic acid during synthesis. “Purified” can also be used to refer to a nucleic acid separated from components that can accompany it during production of the nucleic acid (e.g., during synthesis in vitro, etc.). In some cases, a nucleic acid (or a mixture of nucleic acids) is substantially pure when the nucleic acid (or mixture of nucleic acids) is at least 60% or at least 75% by weight free from organic molecules (e.g., proteins) with which it is naturally associated or with which it is associated during production. In some cases, the nucleic acid is from 30% to 60% pure. In some cases, the nucleic acid (or mixture of nucleic acid) is at least 60%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, by weight, pure.

[0044] The terms “polynucleotide” and “nucleic acid,” used interchangeably herein, refer to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. Thus, this term includes, but is not limited to, single-, double-, or multi-stranded DNA or RNA, genomic DNA, cDNA, DNA-RNA hybrids, or a polymer comprising purine and pyrimidine bases or other natural, chemically or biochemically modified, non-natural, or derivatized nucleotide bases. In some cases, a polynucleotide is RNA. In some cases, a polynucleotide is DNA. A “polynucleotide” includes a nucleic acid that is incorporated into a viral vector or a bacterial vector. A “polynucleoside” is a polymeric form ofnucleosides or any length, and may include chemically or biochemically modified, non-natural, or derivatized nucleosides.

[0045] The terms ''peptide," ''polypeptide," and "protein" are used interchangeably herein, and refer to a polymeric form of amino acids of any length, which can include coded and non-coded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones. The term “polypeptide” includes glycosylated polypeptides.

[0046] The term “heterologous” refers to two components that are defined by structures derived from different sources. For example, where “heterologous” is used in the context of a polypeptide, the polypeptide includes operably linked amino acid sequences that can be derived from one or more different polypeptides, e.g., amino acid sequences that are not operably linked to the polypeptide in nature. As another example, where a composition comprises an HCV E1 / E2 heterodimer and a “heterologous” polypeptide, the “heterologous polypeptide is a polypeptide other than HCV E1 or HCV E2. As another example, where a fusion polypeptide comprises: a) a T-cell epitope polypeptide (a “TP”); and b) a heterologous fusion partner polypeptide, the “heterologous fusion partner polypeptide” is one that is not found associated with the T-cell epitope polypeptide in nature. As another example, where “heterologous” is used in the context of a nucleic acid or nucleotide sequence, the nucleic acid includes operably linked nucleotide sequences that are not normally linked in nature. For example, an IRES can be heterologous to a nucleotide sequence encoding an HCV E1 and / or E2 polypeptide, where the IRES is from an organism (e.g., a virus) other than HCV.

[0047] A "conservative amino acid substitution" is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degree of similarity may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well known to those of skill in the art. See, e.g., Pearson (1994) Methods Mol. Biol.24: 307-331, herein incorporated by reference. Examples of groups of amino acids that have side chains with similar chemical properties, and that thus constitute conservative amino acid substitution groups, include: 1) aliphatic side chain-containing amino acids: glycine, alanine, valine, leucine and isoleucine; 2) aliphatic- hydroxyl side chain-containing amino acids: serine and threonine; 3) amino acids with amide-containing side chains: asparagine and glutamine; 4) aromatic side chain-containing amino acids: phenylalanine, tyrosine, and tryptophan; 5) amino acids with basic side chains: lysine, arginine, and histidine; 6) amino acids with acidic side chains: aspartate and glutamate, and 7) amino acids with sulfur-containing side chains: cysteine and methionine. Examples of conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine.

[0048] Before the present invention is further described, it is to be understood that this invention is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0049] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0050] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[0051] It must be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a nucleic acid” includes a plurality of such nucleic acid and reference to “the nucleoside base modification” includes reference to one or more nucleoside base modifications and equivalents thereof known to those skilled in the art, and so forth. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation.

[0052] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the invention are specifically embraced by the present invention and aredisclosed herein just as if each and every combination was individually and explicitly disclosed. In addition, all sub-combinations of the various embodiments and elements thereof are also specifically embraced by the present invention and are disclosed herein just as if each and every such sub- combination was individually and explicitly disclosed herein.

[0053] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed. DETAILED DESCRIPTION

[0054] The present disclosure provides nucleic acids comprising nucleotide sequences encoding hepatitis C virus (HCV) E1 and / or E2 polypeptides. The nucleic acids can comprise one or more modifications. The nucleic acids can be self-amplifying. The present disclosure provides compositions comprising the nucleic acids. The present disclosure provides methods of inducing an immune response to HCV in an individual, comprising administering to the individual a composition of the present disclosure. NUCLEIC ACID COMPOSITIONS

[0055] The present disclosure provides compositions (e.g., immunogenic compositions) comprising one or more nucleic acids comprising nucleotide sequences encoding one or more of: i) an HCV E1 polypeptide; ii) an HCV E2 polypeptide; iii) an HCV E1 / E2 heterodimer; iv) an HCV core polypeptide; and v) a T-cell epitope polypeptide (a polypeptide other than an HCV E1 or E2 polypeptide; also referred to as a “TP”) as described below. In some cases, the one or more nucleic acids are DNA molecules. In some cases, the one or more nucleic acids are RNA (e.g., mRNA) molecules.

[0056] In some cases, a nucleic acid composition of the present disclosure comprises: a) one or more RNA molecules; and b) one or more of: i) a lipid; ii) a polymer; iii) a peptide; iv) a viral-like particle; and v) a cationic nanoemulsion; where the one or more RNA molecules comprises a polypeptide- encoding region comprising one or more nucleotide sequences encoding an HCV E1 polypeptide and / or an HCV E2 polypeptide. In some cases, a nucleic acid composition of the present disclosure comprises: a) one or more RNA molecules; and b) one or more of: i) a lipid; ii) a polymer; iii) a peptide; iv) a viral- like particle; and v) a cationic nanoemulsion; where the one or more RNA molecules comprises: i) a 5’ untranslated region (5’ UTR), e.g., a 5’ IRES; and ii) a polypeptide-encoding region comprising one or more nucleotide sequences encoding an HCV E1 polypeptide and / or an HCV E2 polypeptide. In some cases, a nucleic acid composition of the present disclosure comprises: a) one or more RNA molecules; and b) one or more of: i) a lipid; ii) a polymer; iii) a peptide; iv) a viral-like particle; and v) a cationicnanoemulsion; where the one or more RNA molecules comprises: i) a 5’ UTR; and ii) a polypeptide- encoding region comprising one or more nucleotide sequences encoding an HCV E1 polypeptide and / or an HCV E2 polypeptide. The one or more RNA molecules can further include additional polypeptide- encoding regions, as described below. The present disclosure provides nucleic acids comprising nucleotide sequences encoding HCV E1 and / or E2 polypeptides and / or one or more T-cell epitope polypeptides. In some cases, the one or more RNA molecules are mRNA. In some cases, the one or more RNA molecules are self-replicating RNA molecules.

[0057] In some cases, a nucleic acid composition of the present disclosure comprises: a) one or more RNA molecules; and b) one or more of: i) a lipid; ii) a polymer; iii) a peptide; iv) a viral-like particle; and v) a cationic nanoemulsion; where the one or more RNA molecules comprises: i) at least one 5’ cap structure; and ii) a polypeptide-encoding region comprising one or more nucleotide sequences encoding an HCV E1 polypeptide and / or an HCV E2 polypeptide. In some cases, a nucleic acid composition of the present disclosure comprises: a) one or more RNA molecules; and b) one or more of: i) a lipid; ii) a polymer; iii) a peptide; iv) a viral-like particle; and v) a cationic nanoemulsion; where the one or more RNA molecules comprises: i) at least one 5’ cap structure and / or a 5’ untranslated region (5’ UTR); and ii) a polypeptide-encoding region comprising one or more nucleotide sequences encoding an HCV E1 polypeptide and / or an HCV E2 polypeptide. In some cases, a nucleic acid composition of the present disclosure comprises: a) one or more RNA molecules; and b) one or more of: i) a lipid; ii) a polymer; iii) a peptide; iv) a viral-like particle; and v) a cationic nanoemulsion; where the one or more RNA molecules comprises: i) a 5’ UTR; and ii) a polypeptide-encoding region comprising one or more nucleotide sequences encoding an HCV E1 polypeptide and / or an HCV E2 polypeptide. The one or more RNA molecules can further include additional polypeptide-encoding regions, as described below. The present disclosure provides nucleic acids comprising nucleotide sequences encoding HCV E1 and / or E2 polypeptides and / or one or more T-cell epitope polypeptides. In some cases, the one or more RNA molecules are mRNA. In some cases, the one or more RNA molecules are self-replicating RNA molecules.

[0058] In some cases, a nucleic acid composition of the present disclosure comprises: a) one or more RNA molecules; and b) a polyalkyleneimine; where the one or more RNA molecules comprises: i) at least one 5’ cap structure and / or a 5’ UTR; and a polypeptide-encoding region comprising one or more nucleotide sequences encoding an HCV E1 polypeptide and / or an HCV E2 polypeptide. The one or more RNA molecules can further include additional polypeptide-encoding regions, as described below. In some cases, the one or more RNA molecules are self-replicating RNA molecules. 5’ cap structure

[0059] As noted above, an RNA of the present disclosure in some cases comprises a 5’ cap structure. The 5' cap structure of an mRNA is involved in nuclear export, increasing mRNA stability and binds themRNA Cap Binding Protein (CBP), which is responsible for mRNA stability in the cell and translation competency through the association of CBP with poly(A) binding protein to form the mature cyclic mRNA species. The cap further assists the removal of 5' proximal introns removal during mRNA splicing.

[0060] RNA molecules (e.g., mRNA molecules) may be 5'-end capped, generating a 5'-ppp-5'- triphosphate linkage between a terminal guanosine cap residue and the 5'-terminal transcribed sense nucleotide of the mRNA molecule. This 5'-guanylate cap may then be methylated to generate an N7- methyl-guanylate residue. The ribose sugars of the terminal and / or anteterminal transcribed nucleotides of the 5' end of the mRNA may optionally also be 2'-O-methylated.

[0061] Additional modifications include, but are not limited to, 2'-O-methylation of the ribose sugars of 5'-terminal and / or 5'-anteterminal nucleotides of the mRNA (as mentioned above) on the 2'-hydroxyl group of the sugar ring. Multiple distinct 5'-cap structures can be used to generate the 5'-cap of a nucleic acid molecule, such as an mRNA molecule.

[0062] Cap analogs, which herein are also referred to as synthetic cap analogs, chemical caps, chemical cap analogs, or structural or functional cap analogs, differ from natural (i.e., endogenous, wild-type or physiological) 5'-caps in their chemical structure, while retaining cap function. Cap analogs may be chemically (i.e., non-enzymatically) or enzymatically synthesized and / or linked to a nucleic acid molecule. For example, the Anti-Reverse Cap Analog (ARCA) cap contains two guanines linked by a 5'- 5'-triphosphate group, wherein one guanine contains an N7 methyl group as well as a 3'-O-methyl group (i.e., N7,3'-O-dimethyl-guanosine-5'-triphosphate-5'-guanosine (m.sup.7G-3'mppp-G; which may equivalently be designated 3' O-Me-m7G(5')ppp(5')G). The 3'-O atom of the other, unmodified, guanine becomes linked to the 5'-terminal nucleotide of the capped nucleic acid molecule (e.g., an mRNA or mmRNA). The N7- and 3'-O-methlyated guanine provides the terminal moiety of the capped RNA molecule.

[0063] Another exemplary cap is mCAP, which is similar to ARCA but has a 2'-O-methyl group on guanosine (i.e., N7,2'-O-dimethyl-guanosine-5'-triphosphate-5'-guanosine, m.sup.7Gm-ppp-G).

[0064] While cap analogs allow for the concomitant capping of a nucleic acid molecule in an in vitro transcription reaction, up to 20% of transcripts can remain uncapped. This, as well as the structural differences of a cap analog from an endogenous 5'-cap structures of nucleic acids produced by the endogenous, cellular transcription machinery, may lead to reduced translational competency and reduced cellular stability.

[0065] An RNA molecule of the present disclosure may also be capped post-transcriptionally, using enzymes, in order to generate more authentic 5'-cap structures. As used herein, the phrase "more authentic" refers to a feature that closely mirrors or mimics, either structurally or functionally, anendogenous or wild type feature. That is, a "more authentic" feature is better representative of an endogenous, wild-type, natural or physiological cellular function and / or structure as compared to synthetic features or analogs, etc., of the prior art, or which outperforms the corresponding endogenous, wild-type, natural or physiological feature in one or more respects. Non-limiting examples of more authentic 5'cap structures of the present invention are those which, among other things, have enhanced binding of cap binding proteins, increased half-life, reduced susceptibility to 5' endonucleases and / or reduced 5'decapping, as compared to synthetic 5'cap structures known in the art (or to a wild-type, natural or physiological 5'cap structure). For example, recombinant Vaccinia Virus Capping Enzyme and recombinant 2'-O-methyltransferase enzyme can create a canonical 5'-5'-triphosphate linkage between the 5'-terminal nucleotide of an mRNA and a guanine cap nucleotide wherein the cap guanine contains an N7 methylation and the 5'-terminal nucleotide of the mRNA contains a 2'-O-methyl. Such a structure is termed the Cap1 structure. This cap results in a higher translational-competency and cellular stability and a reduced activation of cellular pro-inflammatory cytokines, as compared, e.g., to other 5'cap analog structures known in the art. Cap structures include, but are not limited to, 7mG(5')ppp(5')N, pN2p (cap 0), 7mG(5')ppp(5')NlmpNp (cap 1), and 7mG(5')-ppp(5')NlmpN2mp (cap 2).

[0066] A 5' terminal cap suitable for inclusion in an RNA molecule of the present disclosure can include endogenous caps or cap analogs. A 5' terminal cap suitable for inclusion in an RNA molecule of the present disclosure can comprise a guanine analog. Useful guanine analogs include, but are not limited to, inosine, N1-methyl-guanosine, 2'fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino- guanosine, LNA-guanosine, and 2-azido-guanosine.

[0067] In some cases, the 5’ cap structure present in an RNA molecule of the present disclosure is selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2'fluoro- guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido- guanosine. 5’ UTR

[0068] An RNA molecule of the present disclosure can include a 5’ UTR. Natural 5'UTRs bear features which play roles in for translation initiation. They harbor signatures like Kozak sequences which are commonly known to be involved in the process by which the ribosome initiates translation of many genes. Kozak sequences have the consensus CCR(A / G)CCAUGG, where R is a purine (adenine or guanine) three bases upstream of the start codon (AUG), which is followed by another `G`.5'UTR also have been known to form secondary structures which are involved in elongation factor binding.

[0069] By engineering the features typically found in abundantly expressed genes of specific target organs, one can enhance the stability and protein production of the polynucleotides, primary constructs or mRNA of the disclosure. For example, introduction of 5' UTR of liver-expressed mRNA, such as albumin, serum amyloid A, Apolipoprotein A / B / E, transferrin, alpha fetoprotein, erythropoietin, orFactor VIII, could be used to enhance expression of an RNA molecule, in hepatic cell lines or liver. Likewise, use of 5' UTR from other tissue-specific mRNA to improve expression in that tissue is possible for muscle (MyoD, Myosin, Myoglobin, Myogenin, Herculin), for endothelial cells (Tie-1, CD36), for myeloid cells (C / EBP, AML1, G-CSF, GM-CSF, CD11b, MSR, Fr-1, i-NOS), for leukocytes (CD45, CD18), for adipose tissue (CD36, GLUT4, ACRP30, adiponectin) and for lung epithelial cells (SP-A / B / C / D).

[0070] Other non-UTR sequences may be incorporated into the 5' (or 3' UTR) UTRs. For example, introns or portions of introns sequences may be incorporated into the flanking regions of an RNA molecule of the present disclosure. Incorporation of intronic sequences may increase protein production as well as mRNA levels.

[0071] The following are non-limiting examples of suitable 5’ UTR sequences. Variants may be used, in which one or more nucleotides are added to, or removed from, the termini.

[0072] 5’UTR #1: GGGAAATAAGAGAGAAAAGA AGAGTAAGAAGAAATATAAG AGCCACC (SEQ ID NO:142);

[0073] 5’UTR #2: GGGAGATCAGAGAGAAAAGA AGAGTAAGAAGAAATATAAG AGCCACC (SEQ ID NO:157);

[0074] 5’UTR #3: GGAATAAAAGTCTCAACACA ACATATACAAAACAAACGAA TCTCAAGCAATCAAGCATTC TACTTCTATTGCAGCAATTT AAATCATTTCTTTTAAAGCA AAAGCAATTTTCTGAAAATT TTCACCATTTACGAACGATA GCAAC (SEQ ID NO:158); and

[0075] 5’UTR #4: GGGAGACAAGCUUGGCAUUCCGGUACUGUUGGUAAAGCCA CC (SEQ ID NO:159).

[0076] In some cases, the 5’ UTR is a 5’ IRES. Many IRES sequences are known in the art; any known IRES is suitable for use. Suitable 5’ IRESs are described below. Non-limiting examples of suitable IRESs are depicted in FIG.15A-15G. 3’ UTR

[0077] An RNA molecule of the present disclosure can include a 3’ untranslated region (3’ UTR).3' UTRs are known to have stretches of Adenosines and Uridines embedded in them. These AU rich signatures are particularly prevalent in genes with high rates of turnover. Based on their sequence features and functional properties, the AU rich elements (AREs) can be separated into three classes: Class I AREs contain several dispersed copies of an AUUUA motif within U-rich regions. C-Myc and MyoD contain class I AREs. Class II AREs possess two or more overlapping UUAUUUA(U / A)(U / A) nonamers. Molecules containing this type of AREs include GM-CSF and TNF-a. Class III ARES are less well defined. These U rich regions do not contain an AUUUA motif. c-Jun and Myogenin are two well- studied examples of this class. Most proteins binding to the AREs are known to destabilize themessenger, whereas members of the ELAV family, most notably HuR, have been documented to increase the stability of mRNA. HuR binds to AREs of all the three classes. Engineering the HuR specific binding sites into the 3' UTR of nucleic acid molecules will lead to HuR binding and thus, stabilization of the message in vivo. Examples of suitable 3’ UTRs include those described in USPN 9,827,332. Poly-A tails

[0078] An RNA molecule of the present disclosure can include a poly(adenine) (poly-A) tail. The poly- A tail can be from 30 nucleotides to 250 nucleotides in length.

[0079] A nucleic acid (e.g., a DNA molecule) comprising a nucleotide sequence encoding RNA of the present disclosure can include a polyadenylation signal. Suitable polyadenylation signals are known in the art; and any such polyadenylation signals can be suitable for use. As is known in the art, a polyadenylation signal can include an AATAAA motif. RNA modifications

[0080] An RNA molecule of the present disclosure can include one or more of: a) a modified nucleoside comprising a modified nucleoside base; b) a modified nucleoside comprising a modified sugar; c) a modified internucleoside linkage; and d) a modified backbone linking two or more nucleosides.

[0081] A nucleoside is a base-sugar combination. The base portion of the nucleoside is normally a heterocyclic base. The two most common classes of such heterocyclic bases are the purines and the pyrimidines. Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides that include a pentofuranosyl sugar, the phosphate group can be linked to the 2', the 3', or the 5' hydroxyl moiety of the sugar. In forming oligonucleotides, the phosphate groups covalently link adjacent nucleosides to one another to form a linear polymeric compound. In turn, the respective ends of this linear polymeric compound can be further joined to form a circular compound, however, linear compounds are suitable. In addition, linear compounds may have internal nucleotide base complementarity and may therefore fold in a manner as to produce a fully or partially double-stranded compound. Within oligonucleotides, the phosphate groups are commonly referred to as forming the internucleoside backbone of the oligonucleotide. The normal linkage or backbone of RNA and DNA is a 3' to 5' phosphodiester linkage.

[0082] Suitable nucleic acid modifications include, but are not limited to: 2’O-methyl modified nucleotides, 2’ Fluoro modified nucleotides, locked nucleic acid (LNA) modified nucleotides, peptide nucleic acid (PNA) modified nucleotides, nucleotides with phosphorothioate linkages, and a 5’ cap (e.g., a 7-methylguanylate cap (m7G)). Additional details and additional modifications are described below.

[0083] A 2'-O-Methyl modified nucleoside (also referred to as 2'-O-Methyl RNA) is a naturally occurring modification of RNA found in tRNA and other small RNAs that arises as a post-transcriptionalmodification. RNA can be directly synthesized so that it contains one or more 2'-O-Methyl nucleosides. Such an RNA is stable with respect to attack by single-stranded ribonucleases.

[0084] 2’ Fluoro modified nucleosides (e.g., 2' Fluoro bases) have a fluorine modified ribose which increases binding affinity (Tm) and also confers some relative nuclease resistance when compared to native RNA.

[0085] LNA bases have a modification to the ribose backbone that locks the base in the C3'-endo position, which favors RNA A-type helix duplex geometry. This modification significantly increases Tm and is also very nuclease resistant. Multiple LNA insertions can be placed in an RNA at any position except the 3'-end.

[0086] The phosphorothioate (PS) bond (i.e., a phosphorothioate linkage) substitutes a sulfur atom for a non-bridging oxygen in the phosphate backbone of a nucleic acid (e.g., an oligo). This modification renders the internucleoside linkage resistant to nuclease degradation. Phosphorothioate bonds can be introduced between the last 3-5 nucleosides at the 5'- or 3'-end of an RNA molecule to inhibit exonuclease degradation. Including phosphorothioate bonds within an RNA molecule can help reduce attack by endonucleases as well.

[0087] In some cases, a subject nucleic acid has one or more nucleotides that are 2'-O-Methyl modified nucleosides. In some cases, a subject nucleic acid has one or more 2’ Fluoro modified nucleosides. In some cases, a subject nucleic acid has one or more LNA bases. In some cases, a subject nucleic acid has one or more nucleosides that are linked by a phosphorothioate bond (i.e., the subject nucleic acid has one or more phosphorothioate linkages). Modified backbones and modified internucleoside linkages

[0088] Examples of suitable nucleic acids (e.g., RNA molecules) containing modifications include nucleic acids containing modified backbones or non-natural internucleoside linkages. Nucleic acids having modified backbones include those that retain a phosphorus atom in the backbone and those that do not have a phosphorus atom in the backbone.

[0089] Suitable modified oligonucleotide backbones containing a phosphorus atom therein include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosphonates, 5'- alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3'-amino phosphoramidate and aminoalkylphosphoramidates, phosphorodiamidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, selenophosphates and boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of these, and those having inverted polarity wherein one or more internucleoside linkages is a 3' to 3', 5' to 5' or 2' to 2' linkage. Suitable RNA molecules having inverted polarity comprise a single 3' to 3' linkage at the 3'-most internucleoside linkage i.e. a singleinverted nucleoside residue which may be a basic (the nucleobase is missing or has a hydroxyl group in place thereof). Various salts (such as, for example, potassium or sodium), mixed salts and free acid forms are also included.

[0090] In some cases, a subject nucleic acid comprises one or more phosphorothioate and / or heteroatom internucleoside linkages, in particular -CH2-NH-O-CH2-, -CH2-N(CH3)-O-CH2- (known as a methylene (methylimino) or MMI backbone), -CH2-O-N(CH3)-CH2-, -CH2-N(CH3)-N(CH3)-CH2- and -O-N(CH3)- CH2-CH2- (wherein the native phosphodiester internucleoside linkage is represented as -O-P(=O)(OH)- O-CH2-). MMI type internucleoside linkages are disclosed in U.S. Pat. No.5,489,677, the disclosure of which is incorporated herein by reference in its entirety. Suitable amide internucleoside linkages are disclosed in U.S. Pat. No.5,602,240, the disclosure of which is incorporated herein by reference in its entirety.

[0091] Also suitable are nucleic acids having morpholino backbone structures as described in, e.g., U.S. Pat. No.5,034,506. For example, in some cases, a subject nucleic acid comprises a 6-membered morpholino ring in place of a ribose ring. In some of these instances, a phosphorodiamidate or other non- phosphodiester internucleoside linkage replaces a phosphodiester linkage.

[0092] Suitable modified polynucleoside backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; riboacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CH2component parts. Mimetics

[0093] A subject nucleic acid can be a nucleic acid mimetic. The term "mimetic" as it is applied to polynucleotides is intended to include polynucleotides wherein only the furanose ring or both the furanose ring and the internucleoside linkage are replaced with non-furanose groups, replacement of only the furanose ring is also referred to in the art as being a sugar surrogate. The heterocyclic base moiety or a modified heterocyclic base moiety is maintained for hybridization with an appropriate target nucleic acid. One such nucleic acid, a polynucleoside mimetic that has been shown to have excellent hybridization properties, is referred to as a peptide nucleic acid (PNA). In PNA, the sugar-backbone of a polynucleoside is replaced with an amide containing backbone, in particular an aminoethylglycine backbone. The nucleotides are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone.

[0094] One polynucleoside mimetic that has been reported to have excellent hybridization properties is a peptide nucleic acid (PNA). The backbone in PNA compounds is two or more linked aminoethylglycine units which gives PNA an amide containing backbone. The heterocyclic base moieties are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone. Representative U.S. patents that describe the preparation of PNA compounds include, but are not limited to: U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, the disclosures of which are incorporated herein by reference in their entirety.

[0095] Another class of polynucleotide mimetic that has been studied is based on linked morpholino units (morpholino nucleic acid) having heterocyclic bases attached to the morpholino ring. A number of linking groups have been reported that link the morpholino monomeric units in a morpholino nucleic acid. One class of linking groups has been selected to give a non-ionic oligomeric compound. The non- ionic morpholino-based oligomeric compounds are less likely to have undesired interactions with cellular proteins. Morpholino-based polynucleotides are non-ionic mimics of oligonucleotides which are less likely to form undesired interactions with cellular proteins (Dwaine A. Braasch and David R. Corey, Biochemistry, 2002, 41(14), 4503-4510). Morpholino-based polynucleotides are disclosed in U.S. Pat. No.5,034,506, the disclosure of which is incorporated herein by reference in its entirety. A variety of compounds within the morpholino class of polynucleotides have been prepared, having a variety of different linking groups joining the monomeric subunits.

[0096] A further class of polynucleoside mimetic is referred to as cyclohexenyl nucleic acids (CeNA). The furanose ring normally present in an RNA molecule is replaced with a cyclohexenyl ring. CeNA DMT protected phosphoramidite monomers have been prepared and used for oligomeric compound synthesis following classical phosphoramidite chemistry. Fully modified CeNA oligomeric compounds and oligonucleotides having specific positions modified with CeNA have been prepared and studied (see Wang et al., J. Am. Chem. Soc., 2000, 122, 8595-8602, the disclosure of which is incorporated herein by reference in its entirety).

[0097] A further modification includes Locked Nucleic Acids (LNAs) in which the 2'-hydroxyl group is linked to the 4' carbon atom of the sugar ring thereby forming a 2'-C,4'-C-oxymethylene linkage thereby forming a bicyclic sugar moiety. The linkage can be a methylene (-CH2-), group bridging the 2' oxygen atom and the 4' carbon atom wherein n is 1 or 2 (Singh et al., Chem. Commun., 1998, 4, 455-456, the disclosure of which is incorporated herein by reference in its entirety). LNA and LNA analogs display very high duplex thermal stabilities with complementary DNA and RNA (Tm=+3 to +10° C), stability towards 3'-exonucleolytic degradation and good solubility properties. Potent and nontoxic antisense oligonucleotides containing LNAs have been described (e.g., Wahlestedt et al., Proc. Natl. Acad. Sci. U.S.A., 2000, 97, 5633-5638, the disclosure of which is incorporated herein by reference in its entirety).

[0098] The synthesis and preparation of the LNA monomers adenine, cytosine, guanine, 5-methyl- cytosine, thymine and uracil, along with their oligomerization, and nucleic acid recognition properties have been described (e.g., Koshkin et al., Tetrahedron, 1998, 54, 3607-3630, the disclosure of which is incorporated herein by reference in its entirety). LNAs and preparation thereof are also described in WO 98 / 39352 and WO 99 / 14226, as well as U.S. applications 20120165514, 20100216983, 20090041809, 20060117410, 20040014959, 20020094555, and 20020086998, the disclosures of which are incorporated herein by reference in their entirety. Modified sugar moieties

[0099] A subject nucleic acid can also include one or more substituted sugar moieties. Suitable polynucleotides comprise a sugar substituent group selected from: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C1to C10alkyl or C2to C10alkenyl and alkynyl. Particularly suitable are O((CH2)nO)mCH3, O(CH2)nOCH3, O(CH2)nNH2, O(CH2)nCH3, O(CH2)nONH2, and O(CH2)nON((CH2)nCH3)2, where n and m are from 1 to about 10. Other suitable polynucleosides comprise a sugar substituent group selected from: C1to C10lower alkyl, substituted lower alkyl, alkenyl, alkynyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ONO2, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an RNA, or a group for improving the pharmacodynamic properties of an RNA, and other substituents having similar properties. A suitable modification includes 2'- methoxyethoxy (2'-O-CH2CH2OCH3, also known as 2'-O-(2-methoxyethyl) or 2'-MOE) (Martin et al., Helv. Chim. Acta, 1995, 78, 486-504, the disclosure of which is incorporated herein by reference in its entirety) i.e., an alkoxyalkoxy group. A further suitable modification includes 2'- dimethylaminooxyethoxy, i.e., a O(CH2)2ON(CH3)2group, also known as 2'-DMAOE, as described in examples hereinbelow, and 2'-dimethylaminoethoxyethoxy (also known in the art as 2'-O-dimethyl- amino-ethoxy-ethyl or 2'-DMAEOE), i.e., 2'-O-CH2-O-CH2-N(CH3)2.

[0100] Other suitable sugar substituent groups include methoxy (-O-CH3), aminopropoxy (--O CH2CH2CH2NH2), allyl (-CH2-CH=CH2), -O-allyl (--O--CH2—CH=CH2) and fluoro (F).2'-sugar substituent groups may be in the arabino (up) position or ribo (down) position. A suitable 2'-arabino modification is 2'-F. Similar modifications may also be made at other positions on the RNA, particularly the 3' position of the sugar on the 3' terminal nucleoside or in 2'-5' linked oligonucleosides and the 5' position of 5' terminal nucleoside. Base modifications and substitutions

[0101] A subject nucleic acid may also include nucleobase (often referred to in the art simply as "base") modifications or substitutions. As used herein, "unmodified" or "natural" nucleobases include thepurine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U). Modified nucleobases include other synthetic and natural nucleobases such as 5-methylcytosine (5- me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2- thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl (-C=C-CH3) uracil and cytosine and other alkynyl derivatives of pyrimidine bases, 6-azo uracil, cytosine and thymine, 5- uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8- substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 2-F-adenine, 2-amino-adenine, 8- azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3- deazaadenine. Further modified nucleobases include tricyclic pyrimidines such as phenoxazine cytidine(1H-pyrimido(5,4-b)(1,4)benzoxazin-2(3H)-one), phenothiazine cytidine (1H-pyrimido(5,4- b)(1,4)benzothiazin-2(3H)-one), G-clamps such as a substituted phenoxazine cytidine (e.g.9-(2- aminoethoxy)-H-pyrimido(5,4-(b) (1,4)benzoxazin-2(3H)-one), carbazole cytidine (2H-pyrimido(4,5- b)indol-2-one), pyridoindole cytidine (H-pyrido(3',2':4,5)pyrrolo(2,3-d)pyrimidin-2-one).

[0102] Heterocyclic base moieties may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. Further nucleobases include those disclosed in U.S. Pat. No.3,687,808, those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, pages 858-859, Kroschwitz, J. I., ed. John Wiley & Sons, 1990, those disclosed by Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613, and those disclosed by Sanghvi, Y. S., Chapter 15, Antisense Research and Applications, pages 289-302, Crooke, S. T. and Lebleu, B., ed., CRC Press, 1993; the disclosures of which are incorporated herein by reference in their entirety. Certain of these nucleobases are useful for increasing the binding affinity of an oligomeric compound. These include 5-substituted pyrimidines, 6- azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2-aminopropyladenine, 5- propynyluracil and 5-propynylcytosine.5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi et al., eds., Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp.276-278; the disclosure of which is incorporated herein by reference in its entirety) and are suitable base substitutions, e.g., when combined with 2'-O-methoxyethyl sugar modifications. RNA comprising nucleosides of Formula I

[0103] In some cases, a subject RNA comprises n number of linked nucleosides of Formula I:

[0104] wherein:

[0105] U is O, S, N(RU)nu, or C(RU)nu, where nu is an integer nu is an integer from 0 to 2 and each RUis, independently, H, halo, or optionally substituted alkyl; is a single or double bond; is a single bond or absent; each of R1', R2', R1'', R2'', R3, R4, and R5is, independently, H, halo, hydroxy, thiol, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, optionally substituted hydroxyalkoxy, optionally substituted amino, azido, optionally substituted aryl, optionally substituted aminoalkyl, or absent; wherein the combination of R3with one or more of R1', R1'', R2', R2'', or R5can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl; wherein the combination of R5with one or more of R1', R1'', R2', or R2''can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl; and wherein the combination of R4and one or more of R1', R1'', R2', R2'', R3, or R5can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl; each of Y1, Y2, and Y3, is, independently, O, S, -NRN1-, optionally substituted alkylene, or optionally substituted heteroalkylene, wherein RN1is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, or absent; each Y4is, independently, H, hydroxy, thiol, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, optionally substituted alkoxyalkoxy, or optionally substituted amino; each Y5is, independently, O, S, optionally substituted alkylene, or optionally substituted heteroalkylene; n is an integer from 1 to 100,000; and B is a nucleobase, wherein the combination of B and R1', the combination of B and R2', the combination of B and R1'', or the combination of B and R2'' can, taken together with the carbons to which they are attached, optionally form a bicyclic group or wherein thecombination of B, R1'', and R3or the combination of B, R2'', and R3can optionally form a tricyclic or tetracyclic group. In some cases, B is not pseudouridine or 5-methyl-cytidine. In some cases, B is pseudouridine. In some cases, B is 5-methyl-cytidine.

[0106] In some cases, U is O or C(RU)nu, wherein nu is an integer from 1 to 2 and each RUis, independently, H, halo, or optionally substituted alkyl; each of R1, R1', R1'', R2, R2', and R2'', if present, is, independently, H, halo, hydroxy, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, optionally substituted amino, azido, optionally substituted aryl, or optionally substituted aminoalkyl; each of R3and R4is, independently, H, halo, hydroxy, optionally substituted alkyl, or optionally substituted alkoxyalkoxy; each of Y1, Y2, and Y3, is, independently, O, S, --NRN1--, optionally substituted alkylene, or optionally substituted heteroalkylene, wherein RN1is H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl; each Y4is, independently, H, hydroxy, thiol, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, or optionally substituted amino; each Y5is, independently, O or optionally substituted alkylene; and n is an integer from 10 to 10,000. In some cases, each of R1, R1', and R1'', if present, is H. In some cases, each of R2, R2', and R2'', if present, is, independently, H, halo, hydroxy, optionally substituted alkoxy, or optionally substituted alkoxyalkoxy. In some cases, each of R2, R2', and R2'', if present, is H. In some cases, each of R1, R1', and R1'', if present, is, independently, H, halo, hydroxy, optionally substituted alkoxy, or optionally substituted alkoxyalkoxy. In some cases, U is O or C(RU)nu,where nu is an integer from 1 to 2 and each RUis, independently, H, halo, or optionally substituted alkyl; each of R1and R2is independently H, halo, hydroxy, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkoxyalkoxy, optionally substituted amino, azido, optionally substituted aryl, or optionally substituted aminoalkyl; each of R3and R4is, independently, H or optionally substituted alkyl; each of Y1, Y2, and Y3is, independently O, S, -NRN1-, optionally substituted alkylene, or optionally substituted heteroalkylene, wherein RN1is H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl; each Y4is, independently Y, hydroxy, thiol, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, or optionally substituted amino; each Y5is independently O or optionally substituted alkylene; and n is an integer from 10 to 10,000.

[0107] In some cases, a subject RNA comprises n number of linked nucleosides of Formula Ia:(Formula Ia)

[0108] where the substituents are defined as for Formula I, above.

[0109] In some cases, a subject RNA comprises n number of linked nucleosides of Formula Ib:(Formula Ib)

[0110] where the substituents are defined as for Formula I, above.

[0111] In some cases, n number of B is of Formula II, III, or IV,Formula II;

[0112] wherein each V7is, independently, O, S, N(RVe)nv, or C(RVe)nv, wherein nv is an integer from 0 to 2 and each RVeis, independently, H, halo, optionally substituted amino acid, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, or optionally substituted alkynyloxy; each R25is, independently, H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl; each of R26aand R26bis, independently, H, optionally substituted acyl, optionally substituted amino acid, optionally substituted carbamoylalkyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, or optionally substituted alkoxy; each R27is, independently, H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, or optionally substituted amino; each R28is, independently, H, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl; and each R29is, independently, H, optionally substituted acyl, optionally substituted amino acid, optionally substituted carbamoylalkyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted hydroxyalkyl, optionally substituted hydroxyalkenyl, optionally substituted alkoxy, or optionally substituted amino. Self-amplifying RNA

[0113] In some cases, an RNA molecule of the present disclosure is a self-amplifying (also referred to herein as “self-replicating”) RNA.

[0114] A self-replicating RNA molecule (also referred to as a “replicon”) can, when delivered to a vertebrate cell even without any proteins, lead to the production of multiple daughter RNAs by transcription from itself (via an antisense copy which it generates from itself). A self-replicating RNA molecule is thus in certain cases, a (+) strand molecule that can be directly translated after delivery to a cell, and this translation provides an RNA-dependent RNA polymerase (RdRP) which then produces both antisense and sense transcripts from the delivered RNA. Thus, the delivered RNA leads to the production of multiple daughter RNAs. These daughter RNAs, as well as co-linear subgenomic transcripts, may be translated themselves to provide in situ expression of an encoded immunogen, or may be transcribed to provide further transcripts with the same sense as the delivered RNA which are translated to provide in situ expression of the immunogen. The overall result of this sequence of transcriptions is a large amplification in the number of the introduced replicon RNAs and so the encoded immunogen becomes a major polypeptide product of the host cells. For example, a self-amplifying RNA can include 5’ and 3’ conservative sequence elements (CSE), nsP1-4 genes, and a coding sequence encoding one or more antigenic polypeptides (e.g., one or more of HCV E1, HCV E2, a TP, a fusion protein comprising two or more TPs, etc.) where the coding sequence is operably linked to a promoter. Following translation in situ (e.g., in a cell in an individual), the nsP1-4 proteins form an RdRP complex that recognizes the 5’ CSE and 3’ CSE and amplifies the antigen-encoding RNAs.

[0115] One suitable system for achieving self-replication is to use an alphavirus-based RNA replicon. These (+) stranded replicons are translated after delivery to a cell to give of a replicase (or replicase-transcriptase). The replicase is translated as a polyprotein which auto cleaves to provide a replication complex which creates genomic (-) strand copies of the (+) strand delivered RNA. These (-) strand transcripts can themselves be transcribed to give further copies of the (+) stranded parent RNA and also to give a subgenomic transcript which encodes the immunogen. Translation of the subgenomic transcript thus leads to in situ expression of the immunogen by the infected cell. Suitable alphavirus replicons can use a replicase from a sindbis virus, a semliki forest virus, an eastern equine encephalitis virus, a Venezuelan equine encephalitis virus, etc. Mutant or wild-type viruses sequences can be used e.g. the attenuated TC83 mutant of VEEV has been used in replicons. Other RNA virus-based RNA replicons can be used, including those of positive-strand RNA viruses and negative-strand RNA viruses.

[0116] In some cases, self-replicating RNA molecule of the present disclosure encodes (i) a RdRP which can transcribe RNA from the self-replicating RNA molecule and (ii) a polypeptide- encoding region comprising nucleotide sequences encoding one or more nucleotide sequences encoding HCV E1 and / or E2 polypeptides and / or one or more T-cell epitope polypeptides. The RdRP of any RNA virus can be used. The RdRP can be an alphavirus replicase e.g., comprising one or more of alphavirus proteins nsP1, nsP2, nsP3 and nsP4. See, e.g., Geall et al. (2012) Proc. Natl. Acad. Sci. USA 109:14604.

[0117] Whereas natural alphavirus genomes encode structural virion proteins in addition to the non-structural replicase polyprotein, in some cases, a self-replicating RNA molecule of the present disclosure does not encode alphavirus structural proteins. Thus, e.g., a particular self-replicating RNA can lead to the production of genomic RNA copies of itself in a cell, but not to the production of RNA- containing virions. The inability to produce these virions means that, unlike a wild-type alphavirus, the self-replicating RNA molecule cannot perpetuate itself in infectious form. The alphavirus structural proteins which are necessary for perpetuation in wild-type viruses are absent from self-replicating RNAs of the present disclosure and their place is taken by gene(s) encoding an immunogen of interest (HCV E1 and / or E2 polypeptides; T-cell epitope polypeptides; etc.), such that the subgenomic transcript encodes the immunogen rather than the structural alphavirus virion proteins.

[0118] In some cases, a self-replicating RNA molecule of the present disclosure comprises two open reading frames. One open reading frame encodes a replicase (e.g., RdRP) e.g., the first, (5’) open reading frame; the other open reading frame encodes HCV E1 and / or E2 polypeptides (and in some cases additional polypeptides such as T-cell epitope polypeptides), e.g., the second, (3') open reading frame. As another example, the first (5’) open reading frame encodes HCV E1 and / or E2 polypeptides (and in some cases additional polypeptides such as T-cell epitope polypeptides); and the second (3’) open reading frame encodes a replicase (e.g., RdRP).

[0119] A self-replicating RNA molecule of the present disclosure can have a 5' sequence which is compatible with the encoded replicase. A self-replicating RNA molecule of the present disclosure can include a 5’ cap structure, where suitable 5’ cap structures are known in the art and are described above. The 5’ nucleotide of a self-replicating RNA molecule of the present disclosure can include a 5’ triphosphate group. A self-replicating RNA molecule of the present disclosure can include a 3’ poly-A tail. A self-replicating RNA molecule of the present disclosure can include a poly-A polymerase recognition sequence (e.g., AAUAAA) near its 3’ end. (in addition to any 5' cap structure) one or more nucleotides having a modified nucleobase. For instance, a self-replicating RNA molecule of the present disclosure can include one or more modified pyrimidine nucleobases, such as pseudouridine and / or 5 methylcytosine residues. In some cases, however, the self-replicating RNA includes no modified nucleobases, and may include no modified nucleotides i.e., all of the nucleotides in the self-replicating RNA are standard A, C, G and U ribonucleotides (except for any 5' cap structure, which may include a 7' methylguanosine). In other instances, a self-replicating RNA of the present disclosure can include a 5' cap comprising a 7' methylguanosine, and the first 1, 2 or 35' ribonucleotides may be methylated at the 2' position of the ribose.

[0120] In some cases, a self-replicating RNA of the present disclosure is a recombinant alphavirus. An alphavirus found in nature is selected from the group consisting of the following: Barmah Forest virus complex (comprising Barmah Forest virus); Eastern equine encephalitis complex(comprising seven antigenic types of Eastern equine encephalitis virus); Middelburg virus complex (comprising Middelburg virus); Ndumu virus complex (comprising Ndumu virus); Semliki Forest virus complex (comprising Bebaru virus, Chikungunya virus, Mayaro virus and its subtype Una virus, O'Nyong Nyong virus, and its subtype Igbo-Ora virus, Ross River virus and its subtypes Bebaru virus, Getah virus, Sagiyama virus, Semliki Forest virus and its subtype Me Tri virus); Venezuelan equine encephalitis complex (comprising Cabassou virus, Everglades virus, Mosso das Pedras virus, Mucambo virus, Paramana virus, Pixuna virus, Rio Negro virus, Trocara virus and its subtype Bijou Bridge virus, Venezuelan equine encephalitis virus); Western equine encephalitis complex (comprising Aura virus, Babanki virus, Kyzylagach virus, Sindbis virus, Ockelbo virus, Whataroa virus, Buggy Creek virus, Fort Morgan virus, Highlands J virus, Western equine encephalitis virus); and some unclassified viruses including Salmon pancreatic disease virus; Sleeping Disease virus; Southern elephant seal virus; Tonate virus. More preferably, the alphavirus is selected from the group consisting of Semliki Forest virus complex (comprising the virus types as indicated above, including Semliki Forest virus), Western equine encephalitis complex (comprising the virus types as indicated above, including Sindbis virus), Eastern equine encephalitis virus (comprising the virus types as indicated above), Venezuelan equine encephalitis complex (comprising the virus types as indicated above, including Venezuelan equine encephalitis virus).

[0121] In some cases, the alphavirus is not an alphavirus found in nature. Typically, an alphavirus not found in nature is a variant or derivative of an alphavirus found in nature, that is distinguished from an alphavirus found in nature by at least one mutation in the nucleotide sequence, i.e., the genomic RNA. The mutation in the nucleotide sequence may be selected from an insertion, a substitution or a deletion of one or more nucleotides, compared to an alphavirus found in nature. A mutation in the nucleotide sequence may or may not be associated with a mutation in a polypeptide or protein encoded by the nucleotide sequence. For example, an alphavirus not found in nature may be an attenuated alphavirus. An attenuated alphavirus not found in nature is an alphavirus that typically has at least one mutation in its nucleotide sequence by which it is distinguished from an alphavirus found in nature, and that is either not infectious at all, or that is infectious but has a lower disease-producing ability or no disease-producing ability at all. IRES

[0122] In some cases, a nucleic acid of the present disclosure includes one or more (e.g., 1, 2, 3, 4, or 5, or more) internal ribosome entry sites (IRES). IRES elements are able to bypass the ribosome scanning model of 5' methylated cap-dependent translation and begin translation at internal sites. Thus, an IRES provides a structure to which a ribosome can bind that does not need to be at the 5' end of the mRNA. An IRES can therefore allow a ribosome to initiate translation at a second (or additional)initiation codon within a mRNA, allowing more than one polypeptide to be produced from a single mRNA.

[0123] Suitable IRESs include, but are not limited to, a picornavirus IRES, an Aphthovirus IRES, a Kaposi’s sarcoma-associated herpesvirus IRES, a hepatitis A virus IRES, a hepatitis C virus IRES, a pestivirus IRES, a cripavirus IRES, a Friend murine leukemia virus IRES, a Moloney murine leukemia virus IRES, a Rouse sarcoma virus IRES, a human immunodeficiency virus IRES, a mammalian IRES, and the like. In some cases, the IRES is an IRES from a picornavirus (e.g. a pest viruses (CFFV), a polio virus (PV), an encephalomyocarditis virus (ECMV), a foot-and-mouth disease virus (FMDV), a hepatitis C virus (HCV), a classical swine fever virus (CSFV), a murine leukemia virus (MLV), a simian immune deficiency virus (SIV), or a cricket paralysis viruses (CPV). An IRES can be derived from a eukaryotic non-translated region, e.g., from BiP, fibroblast growth factor-1 (FGF-1), FGF-2, and the like.

[0124] In some cases, an RNA molecule of the present disclosure includes a single IRES. In some cases, an RNA molecule of the present disclosure includes more than one IRES. For example, in some cases, an RNA molecule of the present disclosure includes 2, 3, 4, or 5 IRESs. Where an RNA molecule of the present disclosure includes 2 or more IRESs, in some cases, the 2 or more IRESs are the same (have the same sequence). Where an RNA molecule of the present disclosure includes 2 or more IRESs, in some cases, the 2 or more IRESs have different sequences.

[0125] Non-limiting examples of suitable IRESs are depicted in FIG.15A-15G. Self-cleaving peptides

[0126] In some cases, instead of an IRES, an RNA molecule of the present disclosure comprises a nucleotide sequence encoding a self-cleaving peptide. A “self-cleaving peptide” refers to a peptide that functions such that when the polypeptide comprising the proteins and the self-cleaving peptide is produced, it is immediately “cleaved” or separated into distinct and discrete first and second polypeptides without the need for any external cleavage activity. In some cases, a self-cleaving peptide has a length of from about 15 amino acids to about 40 amino acids. In some cases, a self-cleaving peptide has a length of from about 15 amino acids to about 22 amino acids (e.g., 15, 16, 17, 18, 19, 20, 21, or 22 amino acids). In some cases, a self-cleaving peptide has a length of from about 20 amino acids to about 40 amino acids (e.g., from about 20 amino acids to about 25 amino acids, from about 25 amino acids to about 30 amino acids, from about 30 amino acids to about 35 amino acids, or from about 35 amino acids to about 40 amino acids).

[0127] In some cases, a self-cleaving peptide is a 2A self-cleaving peptide from an aphtho- or a cardiovirus. The primary 2A / 2B cleavage of the aptho- and cardioviruses is mediated by 2A “cleaving” at its own C-terminus. In apthoviruses, such as foot-and-mouth disease viruses (FM DV) and equinerhinitis A virus, the 2A peptide is a short section of about 18 amino acids, which, together with the N- terminal residue of protein 2B (a conserved proline residue) represents an autonomous element capable of mediating “cleavage” at its own C-terminus. Also suitable for use is a “2A-like” peptide.

[0128] Suitable “2A-like” self-cleaving peptides include, e.g.: YHADYYKQRLIHDVEMNPGP (SEQ ID NO:160); HYAGYFADLLIHDIETNPGP (SEQ ID NO:161); QCTNYALLKLAGDVESNPGP (SEQ ID NO:162); ATNFSLLKQAGDVEENPGP (SEQ ID NO:163); AARQMLLLLSGDVETNPGP (SEQ ID NO:164); RAEGRGSLLTCGDVEENPGP (SEQ ID NO:165); TRAEIEDELIRAGIESNPGP (SEQ ID NO:166); TRAEIEDELIRADIESNPGP (SEQ ID NO:167); AKFQIDKILISGDVELNPGP (SEQ ID NO:168); SSIIRTKMLVSGDVEENPGP (SEQ ID NO:169); CDAQRQKLLLSGDIEQNPGP (SEQ ID NO:170); EGRGSLLTCGDVEENPGP (SEQ ID NO:171); and YPIDFGGFLVKADSEFNPGP (SEQ ID NO:172).

[0129] Other suitable self-cleaving peptides include: LLNFDLLKLAGDVESNPGP (SEQ ID NO:173); TLNFDLLKLAGDVESNPGP (SEQ ID NO:174); LLKLAGDVESNPGP (SEQ ID NO:175); NFDLLKLAGDVESNPGP (SEQ ID NO:176); QLLNFDLLKLAGDVESNPGP (SEQ ID NO:177); APVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO:178); VTELLYRMKRAETYCPRPLLAIHPTEARHKQKIVAPVKQT (SEQ ID NO:179); LNFDLLKLAGDVESNPGP (SEQ ID NO:180); LLAIHPTEARHKQKIVAPVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO:181); EARHKQKIVAPVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO:182); and VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO:183). Cleavable linkers

[0130] In some cases, in place of an IRES is a nucleotide sequence encoding a cleavable linker, e.g., a proteolytically cleavable linker.

[0131] The proteolytically cleavable linker can include a protease recognition sequence recognized by a protease selected from the group consisting of alanine carboxypeptidase, Armillaria mellea astacin, bacterial leucyl aminopeptidase, cancer procoagulant, cathepsin B, clostripain, cytosol alanyl aminopeptidase, elastase, endoproteinase Arg-C, enterokinase, gastricsin, gelatinase, Gly-X carboxypeptidase, glycyl endopeptidase, human rhinovirus 3C protease, hypodermin C, IgA-specific serine endopeptidase, leucyl aminopeptidase, leucyl endopeptidase, lysC, lysosomal pro-X carboxypeptidase, lysyl aminopeptidase, methionyl aminopeptidase, myxobacter, nardilysin, pancreatic endopeptidase E, picornain 2A, picornain 3C, proendopeptidase, prolyl aminopeptidase, proprotein convertase I, proprotein convertase II, russellysin, saccharopepsin, semenogelase, T-plasminogen activator, thrombin, tissue kallikrein, tobacco etch virus (TEV), togavirin, tryptophanyl aminopeptidase, U-plasminogen activator, V8, venombin A, venombin AB, and Xaa-pro aminopeptidase.

[0132] For example, the proteolytically cleavable linker can comprise a matrix metalloproteinase cleavage site, e.g., a cleavage site for a MMP selected from collagenase-1, -2, and -3 (MMP-1, -8, and -13), gelatinase A and B (MMP-2 and -9), stromelysin 1, 2, and 3 (MMP-3, -10, and - 11), matrilysin (MMP-7), and membrane metalloproteinases (MT1-MMP and MT2-MMP). For example, the cleavage sequence of MMP-9 is Pro-X-X-Hy (SEQ ID NO:184) (wherein, X represents an arbitrary residue; Hy, a hydrophobic residue), e.g., Pro-X-X-Hy-(Ser / Thr) (SEQ ID NO:185), e.g., Pro-Leu / Gln- Gly-Met-Thr-Ser (SEQ ID NO:186) or Pro-Leu / Gln-Gly-Met-Thr (SEQ ID NO:187). Another example of a protease cleavage site is a plasminogen activator cleavage site, e.g., a uPA or a tissue plasminogen activator (tPA) cleavage site. In some cases, the cleavage site is a furin cleavage site. Specific examples of cleavage sequences of uPA and tPA include sequences comprising Val-Gly-Arg. Another example of a protease cleavage site that can be included in a proteolytically cleavable linker is a tobacco etch virus (TEV) protease cleavage site, e.g., ENLYTQS (SEQ ID NO:188), where the protease cleaves between the glutamine and the serine. Another example of a protease cleavage site that can be included in a proteolytically cleavable linker is an enterokinase cleavage site, e.g., DDDDK (SEQ ID NO:189), where cleavage occurs after the lysine residue. Another example of a protease cleavage site that can be included in a proteolytically cleavable linker is a thrombin cleavage site, e.g., LVPR (SEQ ID NO:190). Additional suitable linkers comprising protease cleavage sites include linkers comprising one or more of the following amino acid sequences: LEVLFQGP (SEQ ID NO:191), cleaved by PreScission protease (a fusion protein comprising human rhinovirus 3C protease and glutathione-S-transferase; Walker et al. (1994) Biotechnol.12:601); a thrombin cleavage site, e.g., CGLVPAGSGP (SEQ ID NO:192); SLLKSRMVPNFN (SEQ ID NO:193) or SLLIARRMPNFN (SEQ ID NO:194), cleaved by cathepsin B; SKLVQASASGVN (SEQ ID NO:195) or SSYLKASDAPDN (SEQ ID NO:196), cleaved by an Epstein- Barr virus protease; RPKPQQFFGLMN (SEQ ID NO:197) cleaved by MMP-3 (stromelysin); SLRPLALWRSFN (SEQ ID NO:198) cleaved by MMP-7 (matrilysin); SPQGIAGQRNFN (SEQ ID NO:199) cleaved by MMP-9; DVDERDVRGFASFL SEQ ID NO:200) cleaved by a thermolysin-like MMP; SLPLGLWAPNFN (SEQ ID NO:201) cleaved by matrix metalloproteinase 2(MMP-2); SLLIFRSWANFN (SEQ ID NO:202) cleaved by cathespin L; SGVVIATVIVIT (SEQ ID NO:203) cleaved by cathepsin D; SLGPQGIWGQFN (SEQ ID NO:204) cleaved by matrix metalloproteinase 1 (MMP-1); KKSPGRVVGGSV (SEQ ID NO:205) cleaved by urokinase-type plasminogen activator; PQGLLGAPGILG (SEQ ID NO:206) cleaved by membrane type 1 matrixmetalloproteinase (MT- MMP); HGPEGLRVGFYESDVMGRGHARLVHVEEPHT (SEQ ID NO:207) cleaved by stromelysin 3 (or MMP-11), thermolysin, fibroblast collagenase and stromelysin-1; GPQGLAGQRGIV (SEQ ID NO:208) cleaved by matrix metalloproteinase 13 (collagenase-3); GGSGQRGRKALE (SEQ ID NO:209) cleaved by tissue-type plasminogen activator(tPA); SLSALLSSDIFN (SEQ ID NO:210) cleaved by human prostate-specific antigen; SLPRFKIIGGFN (SEQ ID NO:211) cleaved by kallikrein(hK3); SLLGIAVPGNFN (SEQ ID NO:212) cleaved by neutrophil elastase; and FFKNIVTPRTPP (SEQ ID NO:213) cleaved by calpain (calcium activated neutral protease). Signal sequences

[0133] An mRNA of the present disclosure can include a nucleotide sequence encoding a peptide that facilitates trafficking of an operably linked polypeptide. As used herein, a "signal sequence" or "signal peptide" is a polynucleotide or polypeptide, respectively, which is from about 9 to 200 nucleotides in length (encoding a polypeptide of from about 3 amino acids to about 60 amino acids in length; e.g., from about 15 amino acids to about 25 amino acids in length), which is incorporated at the 5' (or N-terminus) of the coding region or polypeptide encoded, respectively. In some cases, addition of these sequences results in trafficking of the encoded polypeptide to the endoplasmic reticulum through one or more secretory pathways. In some cases, addition of these sequences results in secretion of the encoded polypeptide from the cell. Some signal peptides are cleaved from the protein by signal peptidase after the proteins are transported.

[0134] Many signal sequences are known in the art; any known signal sequence can be used herein. The following are non-limiting examples of suitable signal sequences.

[0135] ATGGACGCAATGAAACGGGGTCTGTGCTGCGTACTCTTGCTTTGTGGAGCTG TGTTCGTGTCACCTTCT (SEQ ID NO:214) (encoding tissue-type plasminogen activator (t-PA) signal peptide MDAMKRGLCCVLLLCGAVFVSPS (SEQ ID NO:63);

[0136] ATGATGCCATCCTCAGTC TCATGGGGTATTTTGCTC TTGGCGGGTCTGTGCTGT CTCGTGCCGGTGTCGCTC GCA (SEQ ID NO:215) (encoding α-1- anti-trypsin signal peptide MMPSSVS WGILLAGL CCLVPVSLA (SEQ ID NO:216); and

[0137] ATGGCGACGCCGCTGCCT CCGCCCTCCCCGCGGCAC CTGCGGCTGCTGCGGCTG CTGCTCTCCGCCCTCGTC CTCGGC (SEQ ID NO:217) (encoding: MATPLPPP SPRHLRLL RLLLSG (SEQ ID NO:218). HCV E1 / E2 heterodimers; HCV E2 polypeptides; HCV E1 polypeptides

[0138] An RNA molecule of the present disclosure can comprise: i) a nucleotide sequence encoding an HCV E1 polypeptide, and not an HCV E2 polypeptide; or ii) a nucleotide sequence encoding an HCV E2 polypeptide, and not an HCV E1 polypeptide; or iii) a first nucleotide sequence encoding an HCV E1 polypeptide; and a second nucleotide sequence encoding an HCV E2 polypeptide; or iv) a nucleotide sequence encoding an HCV E1 polypeptide and an HCV E2 polypeptide.

[0139] HCV E1, E2, and E1 / E2 polypeptides that can be encoded by an RNA molecule of the present disclosure include wild-type HCV E1 polypeptides, wild-type HCV E2 polypeptides and wild- type HCV E1 / E2 polypeptides, i.e., polypeptides comprising wild-type amino acid sequences found in natural isolates. HCV E2 polypeptides that can be encoded by an RNA molecule of the presentdisclosure include wild-type E2 polypeptides and variant E2 polypeptides. HCV E1 polypeptides that can be encoded by an RNA molecule of the present disclosure include wild-type E1 polypeptides and variant E1 polypeptides. E2 polypeptides

[0140] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can have a length of from about 200 amino acids (aa) to about 250 aa, from about 250 aa to about 275 aa, from about 275 aa to about 300 aa, from about 300 aa to about 325 aa, from about 325 aa to about 350 aa, or from about 350 aa to about 365 aa. In some cases, an HCV E2 polypeptide that is encoded by an RNA molecule of the present disclosure is an HCV E2 ectodomain polypeptide (e.g., lacking the transmembrane domain of a naturally-occurring E2 polypeptide). In some cases, an HCV E2 polypeptide that is encoded by an RNA molecule of the present disclosure is a full-length HCV E2 polypeptide. In some cases, an HCV E2 polypeptide that is encoded by an RNA molecule of the present disclosure is a soluble HCV E2 polypeptide.

[0141] In FIG.1A-AC, the amino acid sequence of E2 is amino acid 384 to amino acid 746. In FIG.2A-2B, the amino acid sequence of E2 is amino acid 384 to amino acid 751. In FIG.3A-3C, the amino acid sequence of E2 is amino acid 385 to amino acid 754. In FIG.20A-20B, the amino acid sequence of E2 is amino acid 384 to amino acid 750. As used herein, an "E2 polypeptide" includes a precursor E2 protein, including the signal sequence; includes a mature E2 polypeptide which lacks this sequence; and includes an E2 polypeptide with a heterologous signal sequence. An E2 polypeptide can include a C-terminal membrane anchor sequence which occurs at approximately amino acid positions 715-730 and may extend as far as approximately amino acid residue 746 (see, Lin et al., J. Virol. (1994) 68:5063-5073).

[0142] In some cases, a E2 polypeptide that is encoded by an RNA molecule of the present disclosure lacks a portion of its C-terminal region, e.g., from about amino acid 715 to the C-terminus; from about amino acid 625 to the C-terminus; from about amino acid 661 to the C-terminus; from about amino acid 655 to the C-terminus; from about amino acid 500 to the C-terminus, where the amino acid numbering is with reference to the numbering in FIG.1A-1C. See, e.g., U.S. Patent No.6,521,423. An E2 polypeptide that lacks a C-terminal region, e.g., a C-terminal membrane anchor sequence, is also referred to herein as a “soluble” E2 polypeptide. In some cases, a soluble E2 polypeptide comprises amino acids 384-717 of any one of the amino acid sequences depicted in FIG.1A-1C, FIG.2A-2B, FIG. 3A-3C, or FIG.20A-20B. In some cases, a soluble E2 polypeptide comprises amino acids 384-661 of any one of the amino acid sequences depicted in FIG.1A-1C, FIG.2A-2B, FIG.3A-3C, or FIG.20A- 20B; and lacks the C-terminal portion, e.g., from amino acids 662-746, from amino acids 662 to 751, or from amino acids 662 to754, or from amino acids 662 to 750. A non-limiting example of a nucleotidesequence encoding a soluble E2 polypeptide is provided in FIG.25H. See, e.g., Guest et al. (2021) Proc. Natl. Acad. Sci. USA 18:e2015149118; Cao et al. (2019) PLoS Pathogens 15:e1007759.

[0143] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%,at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence of an E2 polypeptide depicted in FIG.1A-1C, FIG.2A-2C, FIG.3A-3C, or FIG.20A-20B. An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, or at least about 75%, amino acid sequence identity to an amino acid sequence of an E2 polypeptide depicted in FIG.1A-1C, FIG.2A-2C, FIG.3A-3C, or FIG.20A-20B.

[0144] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence of an E2 polypeptide depicted in FIG.1A-1C. For example, an E2 polypeptide of genotype 1 that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384-746 of an amino acid sequence depicted in FIG.1A-1C. For example, an E2 polypeptide of genotype 1A that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384-746 of an amino acid sequence identified as 1A and depicted in FIG.1A-1C. For example, an E2 polypeptide of genotype 1B that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity toamino acids 384-746 of an amino acid sequence identified as 1B and depicted in FIG.1A-1C. For example, an E2 polypeptide of genotype 1C that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384-746 of an amino acid sequence identified as 1C and depicted in FIG.1A-1C.

[0145] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence of an E2 polypeptide depicted in FIG.2A-2C. For example, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384-751 of an amino acid sequence depicted in FIG. 2A-2C. For example, an E2 polypeptide of genotype 2A that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384-751 of the “consensus” amino acid sequence depicted in FIG.2A-2C. For example, an E2 polypeptide of genotype 2B that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384- 751 of the “consensus” amino acid sequence depicted in FIG.2A-2C.

[0146] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acidsequence of an E2 polypeptide depicted in FIG.3A-3C. For example, an E2 polypeptide of genotype 3 that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 385-754 of an amino acid sequence depicted in FIG.3A-3C. For example, an E2 polypeptide of genotype 3A that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 385-754 of an amino acid sequence identified as 3A and depicted in FIG.3A-3C. For example, an E2 polypeptide of genotype 3B that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 385-754 of the amino acid sequence identified as 3B and depicted in FIG.3A-3C. For example, an E2 polypeptide of genotype 3K that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 385-754 of the amino acid sequence identified as 3K and depicted in FIG.3A-3C.

[0147] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the amino acid sequence of the E2 polypeptide depicted in FIG.20A-20B. For example, an E2 polypeptide of genotype 7A that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about99%, or 100%, amino acid sequence identity to amino acids 384-750 of the amino acid sequence depicted in FIG.20A-20B.

[0148] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the amino acid sequence of a genotype 4 E2 polypeptide depicted in FIG.26B, where the E2 polypeptide amino acid sequence is in bold in FIG. 26B. An example of a nucleotide sequence encoding an E2 polypeptide of genotype 4 is provided in FIG.26A, where the nucleotide sequence encoding the E2 polypeptide is in bold.

[0149] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the amino acid sequence of a genotype 5 E2 polypeptide depicted in FIG.27B, where the E2 polypeptide amino acid sequence is in bold in FIG. 27B. An example of a nucleotide sequence encoding an E2 polypeptide of genotype 5 is provided in FIG.27A, where the nucleotide sequence encoding the E2 polypeptide is in bold.

[0150] An E2 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the amino acid sequence of a genotype 6 E2 polypeptide depicted in FIG.28B, where the E2 polypeptide amino acid sequence in bold in FIG.28B. An example of a nucleotide sequence encoding an E2 polypeptide of genotype 6 is provided in FIG.28A, where the nucleotide sequence encoding the E2 polypeptide is in bold. Genotype 1 – H77

[0151] In some cases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 1 E2 polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384 to 746 of the amino acid sequence identified in FIG.1A-1C as “NP_671491 (H77)” (also referred to herein as “H77”); and can have a length of about 363 amino acids (e.g., from 360 amino acids to 365amino acids); where such a polypeptide is referred to as an “H77 E2 polypeptide.” In some cases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384 to 661 of the H77 amino acid sequence depicted in FIG.1A-1C; and can have a length of about 278 amino acids (e.g., from about 275 amino acids to about 280 amino acids), where such an E2 polypeptide is referred to as a “soluble” H77 E2 polypeptide. In some cases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384 to 717 of the H77 amino acid sequence depicted in FIG.1A-1C; and can have a length of about 334 amino acids (e.g., from about 330 amino acids to about 340 amino acids), where such an E2 polypeptide is referred to as a “soluble” H77 E2 polypeptide. Genotype 2 – J6

[0152] In some cases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 2 E2 polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384 to 751 of the amino acid sequence identified in FIG.2A-2B as “D00944 (HC-J6)” (also referred to herein as “J6”); and can have a length of about 368 amino acids (e.g., from 365 amino acids to 370 amino acids); where such a polypeptide is referred to as an “J6 E2 polypeptide.” In some cases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384 to 661 of the J6 amino acid sequence depicted in FIG.2A-2B; and can have a length of about 278 amino acids (e.g., from about 275 amino acids to about 280 amino acids), where such an E2 polypeptide is referred to as a “soluble” J6 E2 polypeptide. In some cases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 384 to 717 of the J6 amino acid sequence depicted in FIG.2A-2B; and can have a length of about 334 amino acids (e.g., from about 330 amino acids to about 340 amino acids), where such an E2 polypeptide is referred to as a “soluble” J6 E2 polypeptide. Genotype 3 – S52

[0153] In some cases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 3 E2 polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 385 to 750 of the amino acid sequence identified in FIG.3A-3C as “ADF96232 (S52)” (also referred to herein as “S52”); and can have a length of about 366 amino acids (e.g., from about 363 amino acids to about 370 amino acids; where such a polypeptide is referred to as an “S52 E2 polypeptide.”. In somecases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 385 to 661 of the S52 amino acid sequence depicted in FIG.3A- 3C; and can have a length of about 278 amino acids (e.g., from about 275 amino acids to about 280 amino acids), where such an E2 polypeptide is referred to as a “soluble” S52 E2 polypeptide. In some cases, an E2 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 385 to 717 of the S52 amino acid sequence depicted in FIG.3A- 3C; and can have a length of about 334 amino acids (e.g., from about 330 amino acids to about 340 amino acids), where such an E2 polypeptide is referred to as a “soluble” S52 E2 polypeptide. E1 polypeptides

[0154] An HCV E1 polypeptide that is encoded by an RNA molecule of the present disclosure can have a length of from about 100 amino acids (aa) to about 150 aa, from about 150 aa to about 175 aa, from about 175 aa to about 195 aa, from about 131 aa to about 175 aa, or from about 175 aa to about 193 aa. In some cases, an HCV E1 polypeptide that is encoded by an RNA molecule of the present disclosure is an HCV E1 ectodomain polypeptide. In some cases, an HCV E1 polypeptide that is encoded by an RNA molecule of the present disclosure is a full-length HCV E1 polypeptide.

[0155] In FIG.1A-1C, the amino acid sequence of E1 is amino acid 192 to amino acid 383. In FIG.2A-2C, the amino acid sequence of E1 is amino acid 192 to amino acid 383. In FIG.3A-3C, the amino acid sequence of E1 is amino acid 192 to amino acid 384. In FIG.20A-20B, the amino acid sequence of E1 is amino acid 192 to amino acid 383. Amino acids at around 170 through approximately 191 serve as a signal sequence for E1. As used herein, "E1 polypeptide" includes a precursor E1 protein, including the signal sequence; includes a mature E1 polypeptide which lacks this sequence; and includes an E1 polypeptide with a heterologous signal sequence. An E1 polypeptide can include a C-terminal membrane anchor sequence which occurs at approximately amino acid positions 360-383 (see, e.g., WO 96 / 04301). In some cases, a suitable E1 polypeptide lacks a C-terminal portion that includes a transmembrane region. For example, in some cases, a suitable E1 polypeptide lacks the C-terminal portion from amino acid 330 to amino acid 384, or from amino acid 360 to amino acid 384. E1 polypeptides can be an E1 polypeptide of any genotype, subtype or isolate of HCV. E1 polypeptides of genotype 1, E1 polypeptides of genotype 2, and E1 polypeptides of genotype 3 can be encoded by an RNA molecule of the present disclosure. An E1 polypeptide that lacks a C-terminal region, e.g., a C- terminal membrane anchor sequence, is also referred to herein as a “soluble” E1 polypeptide. A non- limiting example of a nucleotide sequence encoding a soluble E1 polypeptide is provided in FIG.25G. In some cases, a soluble E1 polypeptide comprises amino acids 192-352 of any one of the amino acid sequences depicted in FIG.1A-1C, FIG.2A-2B, FIG.3A-3C, or FIG.20A-20B. In some cases, a solubleE1 polypeptide comprises amino acids 192-367 of any one of the amino acid sequences depicted in FIG. 1A-1C, FIG.2A-2B, FIG.3A-3C, or FIG.20A-20B; and lacks the C-terminal portion, e.g., from amino acids 352 to 383, from amino acids 367 to 383, or from amino acids 360 to 383. A non-limiting example of a nucleotide sequence encoding a soluble E1 polypeptide is provided in FIG.24A. See, e.g., Guest et al. (2021) Proc. Natl. Acad. Sci. USA 18:e2015149118; Cao et al. (2019) PLoS Pathogens 15:e1007759.

[0156] An E1 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence of an E1 polypeptide depicted in FIG.1A-1C, FIG.2A-2C, FIG.3A-3C, or FIG.20A-20B.

[0157] An E1 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence of an E1 polypeptide depicted in FIG.1A-1C. For example, an E1 polypeptide of genotype 1A that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192-383 of an amino acid sequence identified as 1A and depicted in FIG.1A-1C. For example, an E1 polypeptide of genotype 1B that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192-383 of an amino acid sequence identified as 1B and depicted in FIG.1A-1C. For example, an E1 polypeptide of genotype 1C that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192-383 of an amino acid sequence identified as 1C and depicted in FIG.1A-1C.

[0158] An E1 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence of an E1 polypeptide depicted in FIG.2A-2C. For example, an E1 polypeptide of genotype 2A that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192-383 of an amino acid sequence identified as 2A and depicted in FIG.2A-2C. For example, an E1 polypeptide of genotype 2B that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192-383 of an amino acid sequence identified as 2B and depicted in FIG.2A-2C.

[0159] An E1 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the consensus E1 polypeptide amino acid sequence depicted in FIG.3A-3C.

[0160] An E1 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence of an E1 polypeptide depicted in FIG.20A-20B. For example, an E1 polypeptide of genotype 7A that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about99%, or 100%, amino acid sequence identity to amino acids 192-383 of the amino acid sequence depicted in FIG.20A-20B.

[0161] An E1 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the amino acid sequence of a genotype 4 E1 polypeptide depicted in FIG.26B, where the E1 polypeptide amino acid sequence is underlined in FIG.26B. An example of a nucleotide sequence encoding an E1 polypeptide of genotype 4 is provided in FIG.26A, where the nucleotide sequence encoding the E2 polypeptide is underlined.

[0162] An E1 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the amino acid sequence of a genotype 5 E1 polypeptide depicted in FIG.27B, where the E1 polypeptide amino acid sequence is underlined in FIG.27B. An example of a nucleotide sequence encoding an E1 polypeptide of genotype 5 is provided in FIG.27A, where the nucleotide sequence encoding the E2 polypeptide is underlined.

[0163] An E1 polypeptide that is encoded by an RNA molecule of the present disclosure can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the amino acid sequence of a genotype 6 E1 polypeptide depicted in FIG.28B, where the E1 polypeptide amino acid sequence is underlined in FIG.28B. An example of a nucleotide sequence encoding an E1 polypeptide of genotype 6 is provided in FIG.28A, where the nucleotide sequence encoding the E2 polypeptide is underlined. Genotype 1 – H77

[0164] In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 1 E1 polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 383 of the amino acid sequence identified in FIG.1A-1C as “NP_671491 (H77)” (also referred to herein as “H77”); and can have a length of about 192 amino acids (e.g., from 190 amino acids to 195amino acids); where such a polypeptide is referred to as an “H77 E1 polypeptide.” In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 352 of the H77 amino acid sequence depicted in FIG.1A-1C; and can have a length of about 161 amino acids (e.g., from about 158 amino acids to about 165 amino acids), where such an E1 polypeptide is referred to as a “soluble” H77 E1 polypeptide. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 367 of the H77 amino acid sequence depicted in FIG.1A-1C; and can have a length of about 176 amino acids (e.g., from about 174 amino acids to about 180 amino acids), where such an E1 polypeptide is referred to as a “soluble” H77 E1 polypeptide. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 359 of the H77 amino acid sequence depicted in FIG.1A-1C; and can have a length of about 168 amino acids (e.g., from about 165 amino acids to about 172 amino acids), where such an E1 polypeptide is referred to as a “soluble” H77 E1 polypeptide. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 331 of the H77 amino acid sequence depicted in FIG.1A-1C; and can have a length of about 140 amino acids (e.g., from about 135 amino acids to about 145 amino acids), where such an E1 polypeptide is referred to as a “soluble” H77 E1 polypeptide. Genotype 2 – J6

[0165] In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 2 E1 polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 383 of the amino acid sequence identified in FIG.2A-2B as “D00944 (HC-J6)” (also referred to herein as “J6”); and can have a length of about 192 amino acids (e.g., from 190 amino acids to 195 amino acids); where such a polypeptide is referred to as a “J6 E1 polypeptide.”. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 352 of the J6 amino acid sequence depicted in FIG.2A-2B; and can have a length of about 161 amino acids (e.g., from about 158 amino acids to about 165 amino acids), where such an E1 polypeptide is referred to as a “soluble” J6 E1 polypeptide. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acidsequence identity to amino acids 192 to 367 of the J6 amino acid sequence depicted in FIG.2A-2B; and can have a length of about 176 amino acids (e.g., from about 174 amino acids to about 180 amino acids), where such an E1 polypeptide is referred to as a “soluble” J6 E1 polypeptide. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 359 of the J6 amino acid sequence depicted in FIG.2A-2B; and can have a length of about 168 amino acids (e.g., from about 165 amino acids to about 172 amino acids), where such an E1 polypeptide is referred to as a “soluble” J6 E1 polypeptide. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 331 of the J6 amino acid sequence depicted in FIG.2A-2B; and can have a length of about 140 amino acids (e.g., from about 135 amino acids to about 145 amino acids), where such an E1 polypeptide is referred to as a “soluble” J6 E1 polypeptide. Genotype 3 – S52

[0166] In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 3 E1 polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 384 of the amino acid sequence identified in FIG.3A-3C as “ADF96232 (S52)” (also referred to herein as “S52”); and can have a length of about 193 amino acids (e.g., from 190 amino acids to 196 amino acids); where such a polypeptide is referred to as an “S52 E1 polypeptide.” In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 352 of the S52 amino acid sequence depicted in FIG.3A-3C; and can have a length of about 161 amino acids (e.g., from about 158 amino acids to about 165 amino acids), where such an E1 polypeptide is referred to as a “soluble” S52 E1 polypeptide. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 367 of the S52 amino acid sequence depicted in FIG.3A-3C; and can have a length of about 176 amino acids (e.g., from about 174 amino acids to about 180 amino acids), where such an E1 polypeptide is referred to as a “soluble” S52 E1 polypeptide. In some cases, an E1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 359 of the S52 amino acid sequence depicted in FIG.3A-3C; and can have a length of about 168 amino acids (e.g., from about 165 amino acids to about 172 amino acids), where such an E1 polypeptide is referred to as a “soluble” S52 E1 polypeptide. In some cases, anE1 polypeptide that is encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 192 to 331 of the S52 amino acid sequence depicted in FIG.3A-3C; and can have a length of about 140 amino acids (e.g., from about 135 amino acids to about 145 amino acids), where such an E1 polypeptide is referred to as a “soluble” S52 E1 polypeptide. HCV Core

[0167] In some cases, the polypeptide-encoding region of an RNA of the present disclosure comprises a nucleotide sequence encoding HCV core protein. In some cases, the polypeptide-encoding region encodes a core-E1-E2 polyprotein, which is then processed intracellularly to produce core, E1, and E2 (where the E1 and E2 polypeptides can form a heterodimer). Inclusion of an HCV core protein- encoding nucleotide sequence may result in formation of a viral-like particle (VLP) comprising core, E1, and E2.

[0168] HCV core protein has 191 amino acids and can be divided into three domains on the basis of hydrophobicity: domain 1 (residues 1–117) contains mainly basic residues with two short hydrophobic regions; domain 2 (residues 118–174) is less basic and more hydrophobic and its C- terminus is at the end of p21; domain 3 (residues 175–191) is highly hydrophobic and acts as a signal sequence for E1 envelope protein.

[0169] Amino acid sequences of HCV core proteins of various genotypes are known in the art. See, e.g., the amino acid sequences depicted in FIG.1A-1C, FIG.2A-2C, FIG.3A-3C, FIG.20A-20B, FIG.26B, FIG.27B, and FIG.28B. In some cases, an HCV core protein comprises an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 98%, or 100%, amino acid sequence identity to amino acids 1- 191 of any one of the core-E1-E2 amino acid sequences depicted in FIG.1A-1C, FIG.2A-2C, FIG.3A- 3C, FIG.20A-20B, FIG.26B, FIG.27B, and FIG.28B. In some cases, an HCV core protein has a length of from about 170 amino acids to about 191 amino acids; e.g., in some cases, an HCV core protein has a length of from about 170 amino acids to about 175 amino acids, from about 175 amino acids to about 180 amino acids, from about 180 amino acids to about 185 amino acids, or from about 185 to about 191 amino acids.

[0170] The core protein can be of any HCV genotype. For example, the core protein can be a genotype 1a core protein, a genotype 1b core protein, a genotype 2 core protein, a genotype 3 core protein, a genotype 4 core protein, a genotype 5 core protein, a genotype 6 core protein, etc.

[0171] In some cases, a composition of the present disclosure comprises an RNA molecule comprising: i) a first nucleotide sequence encoding an HCV core polypeptide; ii) a second nucleotide sequence encoding an HCV E1 polypeptide; and iii) a third nucleotide sequence encoding an HCV E2polypeptide. The first, second, and third nucleotide sequence can be separated by IRESs, where the IRESs can be the same or different.

[0172] In some cases, a composition of the present disclosure comprises a nucleotide sequence encoding a core-E1-E2 polyprotein. In some cases, a composition of the present disclosure comprises a nucleotide sequence encoding HCV core protein, a nucleotide sequence encoding an HCV E1 polypeptide, and a nucleotide sequence encoding an HCV E2 polypeptide, where the nucleotide sequence encoding the core protein and the E1 protein are separated by an IRES and where the nucleotide sequence encoding the E1 protein and the E2 protein are separated by an IRES. In some cases, a composition of the present disclosure comprises: i) a first nucleic acid comprising a nucleotide sequence encoding a core-E1-E2 polyprotein; and ii) a second nucleic acid comprising a nucleotide sequence encoding a fusion polypeptide comprising two or more TPs. In some cases, a composition of the present disclosure comprises: i) a first nucleic acid comprising a nucleotide sequence encoding a core-E1-E2 polyprotein; and ii) a second nucleic acid comprising one or more nucleotides encoding one or more TPs.

[0173] For example, as depicted schematically in FIG.5C, in some cases, a composition of the present disclosure comprises: a) a first RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 1a; b) a second RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 2; c) a third RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 3; and d) a fourth RNA molecule comprising one or more nucleotide sequences encoding one or more heterologous polypeptides (e.g., one or more TPs) or a heterologous fusion polypeptide (e.g., a fusion polypeptide comprising two or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 TPs).

[0174] As another example, as depicted schematically in FIG.5D, in some cases, a composition of the present disclosure comprises: a) a first RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 1b; b) a second RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 2; c) a third RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 3; and d) a fourth RNA molecule comprising one or more nucleotide sequences encoding one or more heterologous polypeptides (e.g., one or more TPs) or a heterologous fusion polypeptide (e.g., a fusion polypeptide comprising two or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 TPs).

[0175] As another example, as depicted schematically in FIG.5E, in some cases, a composition of the present disclosure comprises: a) a first RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 1a; b) a second RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 2; c) a third RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 3;d) a fourth RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 1b; b) a second RNA molecule comprising a nucleotide sequence(s) encoding HCV core, HCV E1, and HCVE2 of genotype 2and e) a fifth RNA molecule comprising one or more nucleotide sequences encoding one or more heterologous polypeptides (e.g., one or more TPs) or a heterologous fusion polypeptide (e.g., a fusion polypeptide comprising two or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 TPs).

[0176] In other instances, a separate RNA molecule encodes HCV core protein. For example, in some cases, a composition of the present disclosure comprises: a) a first RNA molecule comprising a nucleotide sequence encoding HCV core; b) a second RNA molecule comprising a nucleotide sequence encoding HCV E1 and / or E2; and c) a third RNA molecule comprising one or more nucleotide sequences encoding one or more heterologous polypeptides (e.g., one or more TPs) or a heterologous fusion polypeptide (e.g., a fusion polypeptide comprising two or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 TPs). Genotype 1 – H77

[0177] In some cases, an HCV Core polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 1 Core polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 1 to 191 of the amino acid sequence identified in FIG.1A-1C as “NP_671491 (H77)” (also referred to herein as “H77”); and can have a length of about 191 amino acids (e.g., from 189 amino acids to 195 amino acids); where such a polypeptide is referred to as an “H77 Core polypeptide.” In some cases, an RNA molecule of the present disclosure comprises a nucleotide sequence encoding a Core-E1-E2 polyprotein. In some cases, the Core-E1-E2 polyprotein comprises: i) an H77 Core polypeptide; ii) an H77 E1 polypeptide, as described above; and iii) an H77 E2 polypeptide, as described above. For example, in some cases, the Core-E1-E2 polyprotein is a genotype 1 Core-E1-E2 polyprotein and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 1 to 746 of the H77 amino acid sequence depicted in FIG.1A-1C. Genotype 2 – J6

[0178] In some cases, an HCV Core polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 2 Core polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 1 to 191 of the amino acid sequence identified in FIG.2A-2B as “D00944 (HC-J6)” (also referred to herein as “J6”); and can have a length of about 191 amino acids (e.g., from 189 amino acids to 195 amino acids); where such a polypeptide is referred to as a “J6 Core polypeptide.” In some cases, an RNA molecule of the present disclosure comprises a nucleotide sequence encoding a Core-E1-E2 polyprotein. In some cases, the Core-E1-E2 polyprotein comprises: i) a J6 Core polypeptide, as described above; andii) a J6 E1 polypeptide; and iii) a J6 E2 polypeptide, as described above. For example, in some cases, the Core-E1-E2 polyprotein is a genotype 2 Core-E1-E2 polyprotein and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 1 to 751 of the J6 amino acid sequence depicted in FIG.2A-2B. Genotype 3 – S52

[0179] In some cases, an HCV Core polypeptide that is encoded by an RNA molecule of the present disclosure is a genotype 3 Core polypeptide and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 1 to 191 of the amino acid sequence identified in FIG.3A-3C as “ADF96232 (S52)” (also referred to herein as “S52”); and can have a length of about 191 amino acids (e.g., from 189 amino acids to 195 amino acids); where such a polypeptide is referred to as an “S52 Core polypeptide.”

[0180] In some cases, an RNA molecule of the present disclosure comprises a nucleotide sequence encoding a Core-E1-E2 polyprotein. In some cases, the Core-E1-E2 polyprotein comprises: i) an S52 Core polypeptide, as described above; and ii) an S526 E1 polypeptide; and iii) an S52 E2 polypeptide, as described above. For example, in some cases, the Core-E1-E2 polyprotein is a genotype 3 Core-E1-E2 polyprotein and comprises an amino acid sequence having at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to amino acids 1 to 750 of the S52 amino acid sequence depicted in FIG.3A-3C. TPs

[0181] In some cases, the polypeptide-encoding region of an RNA of the present disclosure comprises a nucleotide sequence encoding a T-cell epitope polypeptide (a polypeptide other than an HCV E1 or E2 polypeptide) comprising T-cell epitopes (e.g., CD4+and / or CD8+T-cell epitopes that are conserved between or among two or more HCV genotypes and that are presented through one or multiple HLA alleles common within the human population) present in an HCV protein other than E1 and E2. Such T-cell epitope polypeptides are referred to herein as “TPs.”

[0182] Thus, a TP encoded by an RNA molecule of the present disclosure comprises multiple (e.g., 2, 3, 4, or 5, or more than 5) CD4+and / or CD8+T-cell epitopes that are conserved between or among two or more HCV genotypes and that are presented through one or multiple HLA alleles common within the human population. In some cases, a CD8 epitope present in a TP is presented through a single HLA allele (a single HLA haplotype). In some cases, a CD4 epitope present in a TP is presented through multiple different HLA alleles (multiple different HLA haplotypes).

[0183] A TP encoded by an RNA molecule of the present disclosure exhibits one or more of the following features: 1) displays immunodominant epitopes; 2) displays epitopes that are conserved between or among two or more different HCV genotypes; 3) displayed through human leukocyte antigen(HLA) alleles that are common within the human population; 4) good yields by chemical synthesis; and 5) good solubility in a buffered aqueous solution.

[0184] In some cases, a TP encoded by an RNA molecule of the present disclosure displays 1, 2, 3, 4, 5, or from 5 to 10, different epitopes displayed by an HCV polypeptide other than E1 and E2. In some cases, the epitopes are immunodominant epitopes.

[0185] In some cases, a TP encoded by an RNA molecule of the present disclosure displays one or more epitopes that are conserved between or among two or more HCV genotypes. For example, in some cases, a TP suitable for inclusion in a composition of the present disclosure displays 1, 2, 3, 4, 5, or from 5 to 10, epitopes that are conserved between HCV genotype 1 and HCV genotype 3. As another example, in some cases, a TP encoded by an RNA molecule of the present disclosure displays 1, 2, 3, 4, 5, or from 5 to 10, epitopes that are conserved between HCV genotype 1 and HCV genotype 2. As another example, in some cases, a TP encoded by an RNA molecule of the present disclosure displays 1, 2, 3, 4, 5, or from 5 to 10, epitopes that are conserved among HCV genotypes 1, 2, and 3. As another example, in some cases, a TP encoded by an RNA molecule of the present disclosure displays 1, 2, 3, 4, 5, or from 5 to 10, epitopes that are conserved among HCV genotypes 1, 2, 3, and 6. TP35-NS3 T-cell epitope polypeptides

[0186] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least 20% (e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) amino acid sequence identity to the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102; also referred to herein as “TP35-NS3”); where the TP has a length of from 28 amino acids to 35 amino acids (e.g., the T-cell epitope polypeptide has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0187] In some cases, a TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), where X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R; where the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the T-cell epitope polypeptide has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0188] In some cases, a TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KSTKVPVAYAAQGYKVLVLNPSVAATLGFGAYLSK (SEQ ID NO:220); where the TP35-NS3 polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the T-cell epitope polypeptide has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP has a length of 35 amino acids. In some cases, the TTP is part of a fusion polypeptide, as described in detail below.

[0189] In some cases, a TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KSTKVPAAYASQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:221); where the TP35-NS3 T- cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the T-cell epitope polypeptide has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0190] In some cases, a TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KSTKVPAAYVAQGYNVLVLNPSVAATLGFGSFMSR (SEQ ID NO:222); where the TP35-NS3 polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0191] In some cases, a TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSR (SEQ ID NO:223); where the TP35-NS3 polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0192] In some cases, a TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KSTKVPAAYASQGYKVLVLNPSVAATLGFGSYMSK (SEQ ID NO:224); where the TP35-NS3 polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0193] In some cases, a TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence:KSTKVPAAYASQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:221); where the TP35-NS3 polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the T-cell epitope polypeptide has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0194] A TP35-NS3 epitope polypeptide encoded by an RNA molecule of the present disclosure can include a CD8 epitope, e.g., a stretch of 10 contiguous amino acids having the amino acid sequence AYX1X2QGYX3VL (SEQ ID NO:225), where X1is A or V; X2is A or S; and X3is K or N; and / or a stretch of 11 contiguous amino acids having the amino acid sequence ATLGFGX1X2X3SX4(SEQ ID NO:226), where X1is A or S; X2is Y or F; X3is M or L; and X4is K or R. In some cases, the CD8 epitope is a stretch of 10 contiguous amino acids having the amino acid sequence AYAAQGYKVL (SEQ ID NO:227). In some cases, the CD8 epitope is a stretch of 10 contiguous amino acids having the amino acid sequence AYASQGYKVL (SEQ ID NO:228). In some cases, the CD8 epitope is a stretch of 10 contiguous amino acids having the amino acid sequence AYVAQGYNVL (SEQ ID NO:229). A TP35-NS3 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include a CD8 epitope, e.g., a stretch of 10 contiguous amino acids having the amino acid sequence AYAAQGYKVL (SEQ ID NO:227) and / or a stretch of 11 contiguous amino acids having the amino acid sequence ATLGFGAYMSK (SEQ ID NO:230). A TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure can include: i) a stretch of 10 contiguous amino acids having the amino acid sequence AYAAQGYKVL (SEQ ID NO:227); and ii) a stretch of 11 contiguous amino acids having the amino acid sequence ATLGFGAYMSK (SEQ ID NO:330).

[0195] A TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure can include a CD4 epitope, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: VLVLNPSVAATLGFG (SEQ ID NO:231).

[0196] A TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure can include: i) a CD4 epitope, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: VLVLNPSVAATLGFG (SEQ ID NO:231); ii) a first CD8 epitope, e.g., a stretch of 10 contiguous amino acids having the amino acid sequence AYX1X2QGYX3VL (SEQ ID NO:225), where X1is A or V; X2is A or S; and X3is K or N; and iii) a second CD8 epitope, e.g., a stretch of 11 contiguous amino acids having the amino acid sequence ATLGFGX1X2X3SX4(SEQ ID NO:226), where X1is A or S; X2is Y or F; X3is M or L; and X4is K or R. A TP35-NS3 polypeptide encoded by an RNA molecule of the present disclosure can include: i) a CD4 epitope, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: VLVLNPSVAATLGFG (SEQ ID NO:231); ii) a first CD8 epitope, e.g., a stretch of 10 contiguous amino acids having the amino acid sequence AYAAQGYKVL(SEQ ID NO:227); and iii) a second CD8 epitope, e.g., a stretch of 11 contiguous amino acids having the amino acid sequence ATLGFGAYMSK (SEQ ID NO:230).

[0197] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and has a length of from 35 amino acids to 40 amino acids (e.g., 35 amino acids (aa), 36 aa, 37 aa, 38 aa, 39 aa, or 40 aa).

[0198] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and has a length of 35 amino acids. In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequence KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and has a length of 35 amino acids.

[0199] In some cases, a suitable TP is a TP35-NS3 polypeptide with from 1 amino acid to 5 amino acids removed from the N-terminus and / or from 1 amino acid to 5 amino acids removed from the C-terminus. Thus, e.g., a suitable TP can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: PAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:232); and has a length of 30 amino acids. As another example, a suitable TP can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFG (SEQ ID NO:233); and has a length of 30 amino acids.TP50-C T-cell epitope polypeptides

[0200] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least 20% (e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) amino acid sequence identity to the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); also referred to herein as “TP50C”); where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the TP has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0201] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGYP (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K; where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the T-cell epitope polypeptide has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP has a length of 50 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0202] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQ X4IPKX5X6X7X8X9GX10X11WX12X13PGYP (SEQ ID NO:235), where X1is L or V; X2is A; X3is T; X4is P; X5is A or D; X6is R; X7is R or Q; X8is S or P; X9is E or T; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K; where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the T-cell epitope polypeptide has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP has a length of 50 amino acids. In some cases, the T-cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0203] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRPEGRTWAQPGYP (SEQ ID NO:236); where the TTP has a length of from 40 amino acids to 50 amino acids (e.g., the T-cell epitope polypeptide has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP has a length of 50 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0204] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRPEGRAWAQPGYP (SEQ ID NO:237); where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the T-cell epitope polypeptide has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP has a length of 50 amino acids. In some cases, the T-cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0205] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKDRRSTGKSWGKPGYP (SEQ ID NO:238); where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the T-cell epitope polypeptide has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP has a length of 50 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0206] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKDRRSTGKSWGKPGYP (SEQ ID NO:238); where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the TP has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the T-cell epitope polypeptide has a length of 50 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0207] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYVLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:239); where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the T-cell epitope polypeptide has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP has a length of 50 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0208] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRPTGRSWGQPGYP (SEQ ID NO:240); where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the T-cell epitope polypeptide has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa).In some cases, the TP has a length of 50 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0209] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARQPTGRHWAQPGYP (SEQ ID NO:241); where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the TP e has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP has a length of 50 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0210] In some cases, a TP50-C polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GVYLLPRRGPRLGVRTTRKSSERSQPRGRRQRIPKAASSQGKAWGKPGYP (SEQ ID NO:242); where the TP has a length of from 40 amino acids to 50 amino acids (e.g., the TP has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, or 50 aa). In some cases, the TP has a length of 50 amino acids. In some cases, the T-cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0211] A TP50-C polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD4 epitopes, e.g., one or more of: i) a stretch of about 25 contiguous amino acids having the amino acid sequence: RRGPRLGVRATRKTSERSQPRGRRQ (SEQ ID NO:243) or RRGPRLGVRGTRKSSERSQPRGRRQ (SEQ ID NO:244); ii) a stretch of about 24 contiguous amino acids having the amino acid sequence: RATRKTSERSQPRGRRQPIPKARR (SEQ ID NO:245), or RGTRKSSERSQPRGRRQRIPKAAQ (SEQ ID NO:246), or RATRKTSERSQPRGRRQPIPKARQ (SEQ ID NO:247), or RATRKTSERSQPRGRRQPIPKDRR (SEQ ID NO:248); or iii) a stretch of about 20 contiguous amino acids having the amino acid sequence: RRQPIPKARRSEGRSWAQPG (SEQ ID NO:249), or RRQPIPKARPSEGRTWAQPG (SEQ ID NO:250), or RRQPIPKARRPEGRAWAQPG (SEQ ID NO:251), or RRQPIPKDRRSTGKSWGKPG (SEQ ID NO:252), or RRQPIPKDRRSTGKSWGKPG (SEQ ID NO:252), or RRQPIPKARRPTGRSWGQPG (SEQ ID NO:253), or RRQPIPKARQPTGRHWAQPG (SEQ ID NO:254), or RRQRIPKAASSQGKAWGKPG (SEQ ID NO:255).

[0212] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acidsequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and has a length of from 50 amino acids to about 60 amino acids.

[0213] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and has a length of 50 amino acids. In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequence GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and has a length of 50 amino acids. TP23 T-cell epitope polypeptides

[0214] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least 20% (e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) amino acid sequence identity to the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256; also referred to herein as “TP23”); where the TP has a length of from 18 amino acids to 23 amino acids (e.g., the TP has a length of 18 amino acids (aa), 19 aa, 20 aa, 21 aa, 22 aa, or 23 aa). In some cases, the TP has a length of 23 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0215] In some cases, a TP23 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: DVVVX1X2TDALMTGX3TGDFDSVID (SEQ ID NO:257), where X1is V or C; X2is A or S; and X3is F or Y; where the TP23 T-cell epitope polypeptide has a length of from 18 amino acids to 23 amino acids (e.g., the TP has a length of 18 amino acids (aa), 19 aa, 20 aa, 21 aa, 22 aa, or 23 aa). In some cases, the TP has a length of 23 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0216] In some cases, a TP23 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: DVVVVATDALMTGYTGDFDSVID (SEQ ID NO:258); where the TP23 polypeptide has a length of from 18 amino acids to 23 amino acids (e.g., the TP has a length of 18 amino acids (aa), 19 aa, 20 aa, 21 aa, 22 aa, or 23 aa). In some cases, the TP has a length of 23 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0217] In some cases, a TP23 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: DVVVCATDALMTGFTGDFDSVID (SEQ ID NO:259); where the TP23 T-cell epitope polypeptide has a length of from 18 amino acids to 23 amino acids (e.g., T-cell epitope polypeptide has a length of 18 amino acids (aa), 19 aa, 20 aa, 21 aa, 22 aa, or 23 aa). In some cases, the TP has a length of 23 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0218] In some cases, a TP23 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: DVVVCSTDALMTGFTGDFDSVID (SEQ ID NO:260); where the TP23 polypeptide has a length of from 18 amino acids to 23 amino acids (e.g., the TP has a length of 18 amino acids (aa), 19 aa, 20 aa, 21 aa, 22 aa, or 23 aa). In some cases, the TP has a length of 23 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0219] In some cases, a TP23 polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: DVVVCATDALMTGYTGDFDSVID (SEQ ID NO:261); where the TP23 T-cell epitope polypeptide has a length of from 18 amino acids to 23 amino acids (e.g., T-cell epitope polypeptide has a length of 18 amino acids (aa), 19 aa, 20 aa, 21 aa, 22 aa, or 23 aa). In some cases, the TP23 T-cell epitope polypeptide has a length of 23 amino acids. In some cases, the TP23 T-cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0220] A TP23 polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD8 epitopes, e.g., one or both of: i) a stretch of about 9 contiguous amino acids having the amino acid sequence: ATDALMTGF (SEQ ID NO:262), or STDALMTGF (SEQ ID NO:263), or ATDALMTGY (SEQ ID NO:264); or ii) a stretch of about 9 contiguous amino acids having the amino acid sequence GFTGDFDSV (SEQ ID NO:265), or GYTGDFDSV (SEQ ID NO:266).

[0221] A TP23 polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD4 epitopes, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: ATDALMTGFTGDFDS (SEQ ID NO:267), or STDALMTGFTGDFDS (SEQ ID NO:268), or ATDALMTGYTGDFDS (SEQ ID NO:269), or ALMTGFTGDFDSVID (SEQ ID NO:270), or ALMTGYTGDFDSVID (SEQ ID NO:271).

[0222] In some cases, a TP23 polypeptide encoded by an RNA molecule of the present disclosure includes: a) one or more CD8 epitopes, e.g., one or both of: i) a stretch of about 9 contiguous amino acids having the amino acid sequence: ATDALMTGF (SEQ ID NO:262), or STDALMTGF (SEQ ID NO:263), or ATDALMTGY (SEQ ID NO:264); or ii) a stretch of about 9 contiguous amino acids having the amino acid sequence GFTGDFDSV (SEQ ID NO:265), or GYTGDFDSV (SEQ ID NO:266); and b) one or more CD4 epitopes, e.g., a stretch of about 15 contiguous amino acids having the aminoacid sequence: ATDALMTGFTGDFDS (SEQ ID NO:267), or STDALMTGFTGDFDS (SEQ ID NO:268), or ATDALMTGYTGDFDS (SEQ ID NO:269), or ALMTGFTGDFDSVID (SEQ ID NO:270), or ALMTGYTGDFDSVID (SEQ ID NO:271).

[0223] In some cases, an RNA molecule of the present disclosure encodes only a TP comprising an amino acid sequence having at least 20% (e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) amino acid sequence identity to the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256; “TP23”); where the T-cell epitope polypeptide has a length of from 23 amino acids to 30 amino acids (e.g., T-cell epitope polypeptide has a length of 23 amino acids (aa), 24 aa, 25 aa, 26 aa, 27 aa, 28 aa, 29 aa, or 30 aa).

[0224] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and has a length of 23 amino acids. In some cases, a T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the amino acid sequence DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and has a length of 23 amino acids. TP27 T-cell epitope polypeptides

[0225] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least 20% (e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) amino acid sequence identity to the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272; also referred to herein as “TP27”); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0226] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least 20% (e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) amino acid sequence identity to the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLR (SEQ ID NO:103); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP hasa length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the T-cell epitope polypeptide has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0227] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S; where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0228] In some cases, a TP27 T-cell epitope encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: LEVIKGGRHLIFCHSKKKCDELAAKLV (SEQ ID NO:274); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0229] In some cases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: IETIKGGRHLIFCHSKKKCDELAAKLS (SEQ ID NO:275); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0230] In some cases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: LSYIKGGRHLIFCHSKKKCDELAAALR (SEQ ID NO:276); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0231] In some cases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: LAFIKGGRHLIFCHSKKKCDELAALR (SEQ ID NO:277); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0232] In some cases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: IAQLKGGRHLIFCHSKKKCDEIASKLR (SEQ ID NO:278); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0233] In some cases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: LELIKGGRHLIFCHSKKKCDELAQLT (SEQ ID NO:279); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0234] In some cases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: LXLIKGGRHLIFCHSKKKCDELAKQLT (SEQ ID NO:280); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0235] In some cases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: LEYIKGGRHLIFCHSKKKCDELAKQLT (SEQ ID NO:281); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0236] In some cases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: LQHIKGGRHLIFCHSKKKCDELAGKLT (SEQ ID NO:282); where the TP has a length of from 22 amino acids to 27 amino acids (e.g., the TP has a length of 22 amino acids (aa), 23 aa, 24 aa, 25 aa, 26 aa, or 27 aa). In some cases, the TP has a length of 27 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0237] A TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD8 epitopes, e.g., one or both of: i) a stretch of about 9 contiguous amino acids having the amino acid sequence: LIFCHSKKK (SEQ ID NO:283); and ii) a stretch of about 9 contiguous amino acids having the amino acid sequence: HSKKKCDEL (SEQ ID NO:284). In somecases, a TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure includes the following CD8 epitopes: i) a stretch of about 9 contiguous amino acids having the amino acid sequence: LIFCHSKKK (SEQ ID NO:283); and ii) a stretch of about 9 contiguous amino acids having the amino acid sequence: HSKKKCDEL (SEQ ID NO:284).

[0238] A TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD4 epitopes, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: KGGRHLIFCHSKKKCD (SEQ ID NO:285), or a stretch of about 11 contiguous amino acids having the amino acid sequence: GRHLIFCHSKK (SEQ ID NO:286).

[0239] A TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include: a) the following CD8 epitopes: i) a stretch of about 9 contiguous amino acids having the amino acid sequence: LIFCHSKKK (SEQ ID NO:283); and ii) a stretch of about 9 contiguous amino acids having the amino acid sequence: HSKKKCDEL (SEQ ID NO:284); and b) the following CD4 epitope: a stretch of about 11 contiguous amino acids having the amino acid sequence: GRHLIFCHSKK (SEQ ID NO:286). A TP27 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include: a) the following CD8 epitopes: i) a stretch of about 9 contiguous amino acids having the amino acid sequence: LIFCHSKKK (SEQ ID NO:283); and ii) a stretch of about 9 contiguous amino acids having the amino acid sequence: HSKKKCDEL (SEQ ID NO:284); and b) the following CD4 epitope: a stretch of about 15 contiguous amino acids having the amino acid sequence: KGGRHLIFCHSKKKCD (SEQ ID NO:285.

[0240] In some cases, an RNA molecule of the present disclosure encodes only a TP comprising an amino acid sequence having at least 20% (e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) amino acid sequence identity to the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272; “TP27”); where the T-cell epitope polypeptide has a length of from 27 amino acids to 34 amino acids (e.g., T-cell epitope polypeptide has a length of 27 amino acids (aa), 28 aa, 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, or 34 aa).

[0241] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272); and has a length of 27 amino acids. In some cases, a T-cell epitope polypeptide suitable for inclusion in a composition of the presentdisclosure comprises the amino acid sequence LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272); and has a length of 27 amino acids. TP35-NS4 T-cell epitope polypeptides

[0242] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least 20% (e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) amino acid sequence identity to the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287; also referred to herein as “TP35-NS4”); where the TP has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the T-cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0243] In some cases, a TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10(SEQ ID NO:288), where X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; and X10is V or I; where the TP35-NS4 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the T-cell epitope polypeptide has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP35-NS4 T-cell epitope polypeptide has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0244] In some cases, a TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10(SEQ ID NO:289), where X1is I or V; X2is L; X3is R; X4is V or I; X5is P or Q; X6is G or A; X7is A; X8is V or T; X9is S or A; and X10is V; where the TP35-NS4 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP35-NS4 T-cell epitope polypeptide has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0245] In some cases, a TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVAPTHYV (SEQ ID NO:290); where the TP35-NS4 T- cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP35-NS4T-cell epitope polypeptide has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0246] In some cases, a TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGQGEGAVQWMNRLIAFASRGNHVAPTHYV (SEQ ID NO:291); where the TP35-NS4 T- cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP35-NS4 T-cell epitope polypeptide has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0247] In some cases, a TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: VLRRHIGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:292); where the TP35-NS4 T- cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP35-NS4 T-cell epitope polypeptide has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0248] In some cases, a TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPAEGATQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:293); where the TP35-NS4 T- cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP35-NS4 T-cell epitope polypeptide has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0249] In some cases, a TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: IIKRHTGTSEGVTQWMNRLIAFASRGNHVSPTHYI (SEQ ID NO:294); where the TP35-NS4 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids (e.g., the TP has a length of 28 amino acids (aa), 29 aa, 30 aa, 31 aa, 32 aa, 33 aa, 34 aa, or 35 aa). In some cases, the TP35-NS4 T- cell epitope polypeptide has a length of 35 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0250] A TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD8 epitopes, e.g., a stretch of about 16 contiguous amino acids having the amino acid sequence: EGAVQWMNRLIAFASR (SEQ ID NO:295), or EGATQWMNRLIAFASR (SEQ ID NO:296), or EGVTQWMNRLIAFASR (SEQ ID NO:297); or astretch of about 9 contiguous amino acids having the amino acid sequence: WMNRLIAFA (SEQ ID NO:298).

[0251] A TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD4 epitopes, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: RHVGPGEGAVQWMNR (SEQ ID NO:299), or RHVGQGEGAVQWMNR (SEQ ID NO:300), or RHIGPGEGAVQWMNR (SEQ ID NO:301), or RHVGPAEGATQWMNR (SEQ ID NO:302), or RHTGTSEGVTQWMNR (SEQ ID NO:303); or a stretch of about 20 contiguous amino acids having the amino acid sequence: HVGPGEGAVQWMNRLIAFAS (SEQ ID NO:304), or HVGQGEGAVQWMNRLIAFAS (SEQ ID NO:305), or HIGPGEGAVQWMNRLIAFAS (SEQ ID NO:306), or HVGPAEGATQWMNRLIAFAS (SEQ ID NO:307), or HTGTSEGVTQWMNRLIAFAS (SEQ ID NO:308); or a stretch of about 20 contiguous amino acids having the amino acid sequence: GAVQWMNRLIAFASRGNHVS (SEQ ID NO:309), or GAVQWMNRLIAFASRGNHVA (SEQ ID NO:310), or GATQWMNRLIAFASRGNHVS (SEQ ID NO:311), or GVTQWMNRLIAFASRGNHVS (SEQ ID NO:312).

[0252] In some cases, TP35-NS4 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure includes: a) one or more CD8 epitopes, e.g., a stretch of about 16 contiguous amino acids having the amino acid sequence: EGAVQWMNRLIAFASR (SEQ ID NO:295), or EGATQWMNRLIAFASR (SEQ ID NO:296), or EGVTQWMNRLIAFASR (SEQ ID NO:297); or a stretch of about 9 contiguous amino acids having the amino acid sequence: WMNRLIAFA (SEQ ID NO:298); and b) one or more CD4 epitopes, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: RHVGPGEGAVQWMNR (SEQ ID NO:299), or RHVGQGEGAVQWMNR (SEQ ID NO:300), or RHIGPGEGAVQWMNR (SEQ ID NO:301), or RHVGPAEGATQWMNR (SEQ ID NO:302), or RHTGTSEGVTQWMNR (SEQ ID NO:303); or a stretch of about 20 contiguous amino acids having the amino acid sequence: HVGPGEGAVQWMNRLIAFAS (SEQ ID NO:304), or HVGQGEGAVQWMNRLIAFAS (SEQ ID NO:305), or HIGPGEGAVQWMNRLIAFAS (SEQ ID NO:306), or HVGPAEGATQWMNRLIAFAS (SEQ ID NO:307), or HTGTSEGVTQWMNRLIAFAS (SEQ ID NO:308); or a stretch of about 20 contiguous amino acids having amino acid sequence: GAVQWMNRLIAFASRGNHVS (SEQ ID NO:309), or GAVQWMNRLIAFASRGNHVA (SEQ ID NO:310), or GATQWMNRLIAFASRGNHVS (SEQ ID NO:311), or GVTQWMNRLIAFASRGNHVS (SEQ ID NO:312).

[0253] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acidsequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287); and has a length of from 35 amino acids to 40 amino acids (e.g., 35 amino acids (aa), 36 aa, 37 aa, 38 aa, 39 aa, or 40 aa).

[0254] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287); and has a length of 35 amino acids. In some cases, a T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the amino acid sequence ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287); and has a length of 35 amino acids. TP42 T-cell epitope polypeptides

[0255] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG (SEQ ID NO:313); and has a length of from 33 amino acids to about 50 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, 42 aa, or from 42 aa to 50 aa). In some cases, a T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG (SEQ ID NO:313); and has a length of 42 amino acids. In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequence GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG (SEQ ID NO:313); and has a length of 42 amino acids.

[0256] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3GEIPFYGX4AIPX5X6X7X8KGGRHLIFCHSKKKCDEX9AX10X11LX12X13(K)n (SEQ ID NO:314), where X1is G, P, or S; X2is T, N, Q, H, or S; X3is E, T, or D; X4is K or R; X5is L or I; X6is E, A, S, or Q; X7is Q, T, Y, F, or L; X8is I or L; X9is L or I; X10is A, K, or S; X11is K, Q, or A; X12isT, R, or S; and X13is G or S, wherein n is an integer from 2 to 10, and where the TP42 T-cell epitope polypeptide has a length of from 34 amino acids to 52 amino acids (e.g., 34 amino acids (aa), 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, 50 aa, 51 aa, or 52 aa. In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3GEIPFYGX4AIPX5X6X7X8KGGRHLIFCHSKKKCDEX9AX10X11LX12X13KKK (SEQ ID NO:315), where X1is G, P, or S; X2is T, N, Q, H, or S; X3is E, T, or D; X4is K or R; X5is L or I; X6is E, A, S, or Q; X7is Q, T, Y, F, or L; X8is I or L; X9is L or I; X10is A, K, or S; X11is K, Q, or A; X12is T, R, or S; and X13is G or S. In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3GEIPFYGX4AIPX5X6X7X8KGGRHLIFCHSKKKCDEX9AX10X11LX12X13(K)n (SEQ ID NO:352), where X1is G, P, or S; X2is T, N, Q, H, or S; X3is E, T, or D; X4is K or R; X5is L or I; X6is E, A, S, or Q; X7is Q, T, Y, F, V, or L; X8is I or L; X9is L or I; X10is A, K, or S; X11is K, Q, or A; X12is T, R, V, or S; and X13is G or S, wherein n is an integer from 2 to 10, and where the TP42 T-cell epitope polypeptide has a length of from 34 amino acids to 52 amino acids (e.g., 34 amino acids (aa), 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, 48 aa, 49 aa, 50 aa, 51 aa, or 52 aa. In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3GEIPFYGX4AIPX5X6X7X8KGGRHLIFCHSKKKCDEX9AX10X11LX12X13KKK (SEQ ID NO:353), where X1is G, P, or S; X2is T, N, Q, H, or S; X3is E, T, or D; X4is K or R; X5is L or I; X6is E, A, S, or Q; X7is Q, T, Y, F, V, or L; X8is I or L; X9is L or I; X10is A, K, or S; X11is K, Q, or A; X12is T, R, V, or S; and X13is G or S. In some cases, the TP42 T-cell epitope polypeptide has a length of 45 amino acids. In some cases, the TP42 T-cell epitope polypeptide comprises the following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTGKKK (SEQ ID NO:391) and has a length of 45 amino acids.

[0257] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3GEIPFYGX4AIPX5X6X7X8KGGRHLIFCHSKKKCDEX9AX10X11LX12X13(SEQ ID NO:316), where X1is G, P, or S; X2is T, N, Q, H, or S; X3is E, T, or D; X4is K or R; X5is L or I; X6is E, A, S, or Q; X7is Q, T, Y, F, or L; X8is I or L; X9is L or I; X10is A, K, or S; X11is K, Q, or A; X12is T, R, or S; and X13is G or S; where the TP42 T-cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0258] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: STTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVA (SEQ ID NO:317); where the TP42 T-cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0259] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: SNTGEIPFYGKAIPIETIKGGRHLIFCHSKKKCDELAAKLSG (SEQ ID NO:318); where the TP42 T- cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0260] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GQEGEIPFYGRAIPLSYIKGGRHLIFCHSKKKCDELAAALRG (SEQ ID NO:319); where the TP42 T-cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0261] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GHEGEIPFYGKAIPLAFIKGGRHLIFCHSKKKCDELAALRG (SEQ ID NO:320); where the TP42 T- cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0262] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GSEGEIPFYGKAIPIAQLKGGRHLIFCHSKKKCDEIASKLRG (SEQ ID NO:321); where the TP42 T- cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitopepolypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0263] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: PTTGEIPFYGKAIPLELIKGGRHLIFCHSKKKCDELAQLTS (SEQ ID NO:322); where the TP42 T- cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0264] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: PSEGEIPFYGRAIPLXLIKGGRHLIFCHSKKKCDELAKQLTS (SEQ ID NO:323); where the TP42 T- cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0265] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: PTTGEIPFYGKAIPLEYIKGGRHLIFCHSKKKCDELAKQLTS (SEQ ID NO:324); where the TP42 T- cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0266] In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: GNDGEIPFYGKAIPLQHIKGGRHLIFCHSKKKCDELAGKLTS (SEQ ID NO:325); where the TP42 T-cell epitope polypeptide has a length of from 33 amino acids to 42 amino acids (e.g., 33 amino acids (aa), 34 aa, 35 aa, 36 aa, 37 aa, 38 aa, 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, the TP42 T-cell epitope polypeptide has a length of 42 amino acids. In some cases, the TP has a length of 42 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0267] A TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD8 epitopes, e.g., one or more of: i) a stretch of about 11 contiguous amino acids having the amino acid sequence: FYGX1AIPX2X3X4X5(SEQ ID NO:326), where X1is K or R; X2is L or I; X3is E, S, A, Q, or V; X4is Q, T, Y, F, L, or H; and X5is I or L; ii) astretch of about 9 contiguous amino acids having the amino acid sequence: LIFCHSKKK (SEQ ID NO:283); and iii) a stretch of about 9 contiguous amino acids having the amino acid sequence: HSKKKCDEL (SEQ ID NO:284). In some cases, a TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure includes the following CD8 epitopes: i) a stretch of about 11 contiguous amino acids having the amino acid sequence: FYGKAIPLEQI (SEQ ID NO:327); ii) ii) a stretch of about 9 contiguous amino acids having the amino acid sequence: LIFCHSKKK (SEQ ID NO:283); and iii) a stretch of about 9 contiguous amino acids having the amino acid sequence: HSKKKCDEL (SEQ ID NO:284).

[0268] A TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD4 epitopes, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: KGGRHLIFCHSKKKCD (SEQ ID NO:285), or a stretch of about 11 contiguous amino acids having the amino acid sequence: GRHLIFCHSKK (SEQ ID NO:286).

[0269] A TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include: a) the following CD8 epitopes: i) a stretch of about 11 contiguous amino acids having the amino acid sequence: FYGKAIPLEQI (SEQ ID NO:327); ii) ii) a stretch of about 9 contiguous amino acids having the amino acid sequence: LIFCHSKKK (SEQ ID NO:283); and iii) a stretch of about 9 contiguous amino acids having the amino acid sequence: HSKKKCDEL (SEQ ID NO:284); and b) the following CD4 epitope: a stretch of about 11 contiguous amino acids having the amino acid sequence: GRHLIFCHSKK (SEQ ID NO:286). A TP42 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include: a) the following CD8 epitopes: i) a stretch of about 11 contiguous amino acids having the amino acid sequence: FYGKAIPLEQI (SEQ ID NO:327); ii) ii) a stretch of about 9 contiguous amino acids having the amino acid sequence: LIFCHSKKK (SEQ ID NO:283); and iii) a stretch of about 9 contiguous amino acids having the amino acid sequence: HSKKKCDEL (SEQ ID NO:284); and b) the following CD4 epitope: a stretch of about 15 contiguous amino acids having the amino acid sequence: KGGRHLIFCHSKKKCD (SEQ ID NO:285).

[0270] In some cases, a suitable TP is a TP42 polypeptide with from 1 amino acid to 5 amino acids removed from the N-terminus and / or from 1 amino acid to 5 amino acids removed from the C- terminus. Thus, e.g., a suitable TP can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: IPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG (SEQ ID NO:328); and has a length of 37 amino acids. As another example, a suitable TP can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELA (SEQ ID NO:329); and has a length of 37 amino acids. TP45 T-cell epitope polypeptides

[0271] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNV (SEQ ID NO:330); and has a length of from 45 amino acids to about 50 amino acids. In some cases, a T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNV (SEQ ID NO:330); and has a length of 45 amino acids. In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequence LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNV (SEQ ID NO:330); and has a length of 45 amino acids. In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequence LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDKKK (SEQ ID NO:331); and has a length of 45 amino acids.

[0272] In some cases, a TP45 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2AVAX3YRGX4DVX5X6IPX7X8GDVVVX9X10TDALMTGX11TGDFDSVIDX12X13X14(K)n (SEQ ID NO:332), where X1is L or V; X2is N or T; X3is Y or F; X4is L or V; X5is S or A; X6is V or I; X7is T or A; X8is S, Q, or T; X9is V or C; X10is A or S; X11is F or Y; X12is C or K; X13is N or K; and X14is V or K, wherein n is an integer from 2 to 10, and where the TP45 T-cell epitope polypeptide has a length of from 36 amino acids to 55 amino acids. In some cases, a TP45 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2AVAX3YRGX4DVX5X6IPX7X8GDVVVX9X10TDALMTGX11TGDFDSVIDX12X13X14KKK (SEQID NO:333), where X1is L or V; X2is N or T; X3is Y or F; X4is L or V; X5is S or A; X6is V or I; X7is T or A; X8is S, Q, or T; X9is V or C; X10is A or S; X11is F or Y; X12is C or K; X13is N or K; and X14is V or K; where the TP45 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP45 T-cell epitope polypeptide comprises the following amino acid sequence: LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNVKKK (SEQ ID NO:334) and has a length of 48 amino acids.

[0273] In some cases, a TP45 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2AVAX3YRGX4DVX5X6IPX7X8GDVVVX9X10TDALMTGX11TGDFDSVIDX12X13X14(SEQ ID NO:335), where X1is L or V; X2is N or T; X3is Y or F; X4is L or V; X5is S or A; X6is V or I; X7is T or A; X8is S, Q, or T; X9is V or C; X10is A or S; X11is F or Y; X12is C or K; X13is N or K; and X14is V or K; where the TP23 T-cell epitope polypeptide has a length of from 18 amino acids to 23 amino acids (e.g., the TP has a length of 18 amino acids (aa), 19 aa, 20 aa, 21 aa, 22 aa, or 23 aa). In some cases, the TP23 T-cell epitope polypeptide has a length of 23 amino acids. In some cases, the TP23 T- cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0274] In some cases, a TP45 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2AVAX3YRGX4DVX5X6IPX7X8GDVVVX9X10TDALMTGX11TGDFDSVIDX12X13X14(SEQ ID NO:336), where X1is L; X2is N; X3is Y; X4is L; X5is S; X6is V; X7is T; X8is S, Q, or T; X9is V or C; X10is A; X11is F or Y; X12is K; X13is K; and X14is K; where the TP23 T-cell epitope polypeptide has a length of from 18 amino acids to 23 amino acids (e.g., T-cell epitope polypeptide has a length of 18 amino acids (aa), 19 aa, 20 aa, 21 aa, 22 aa, or 23 aa). In some cases, the TP23 T-cell epitope polypeptide has a length of 23 amino acids. In some cases, the TP23 T-cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0275] In some cases, a suitable T-cell epitope polypeptide is a TP45 polypeptide with from 1 amino acid to 5 amino acids removed from the N-terminus and / or from 1 amino acid to 5 amino acids removed from the C-terminus. Thus, for example, in some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVID (SEQ ID NO:337); and has a length of 43 amino acids. For example, in some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequenceLNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVID (SEQ ID NO:337); and has a length of 43 amino acids. TP48 T-cell epitope polypeptides

[0276] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQIL (SEQ ID NO:338); and has a length of from 48 amino acids to about 55 amino acids, or has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQIL (SEQ ID NO:338); and has a length of 48 amino acids. In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequence ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQIL (SEQ ID NO:338); and has a length of 48 amino acids.

[0277] In some cases, a TP48 T-cell epitope encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V, where n is an integer from 2 to 10, and where the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acids. In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19LKKK (SEQ ID NO:340), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T,or G; and X19is I, L, or V, where the TP48 T-cell epitope polypeptide has a length of 51 amino acids. In some cases, the TP48 T-cell epitope polypeptide comprises the amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQILKKK (SEQ ID NO:341) and has a length of 51 amino acids.

[0278] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L (SEQ ID NO:342), where X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V; where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the T-cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0279] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L (SEQ ID NO:343), where X1is I or V; X2is L; X3is R; X4is V or I; X5is P or Q; X6is G or A; X7is A; X8is V or T; X9is S or A; X10is V; X11is P, T, or A; X12is E; X13is S or T; X14is S or A; X15is A, Q, or R; X16is R or K; X17is T; X18is Q, A, or T; and X19is I, L, or V; where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0280] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTAIL (SEQ ID NO:344); where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0281] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids(e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0282] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVAPTHYVTESDASQRVTQLL (SEQ ID NO:346); where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0283] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGQGEGAVQWMNRLIAFASRGNHVAPTHYVAESDASQRVTQVL (SEQ ID NO:347); where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the T- cell epitope polypeptide is part of a fusion polypeptide, as described in detail below.

[0284] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTALL (SEQ ID NO:348); where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0285] In some cases, a TP48 T-cell epitope encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0286] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: VLRRHIGPGEGAVQWMNRLIAFASRGNHVSPTHYV PETDASAKVTQLL (SEQ ID NO:349);where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0287] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPAEGATQWMNRLIAFASRGNHVSPTHYVPETDASRXVTTIL (SEQ ID NO:350); where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0288] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: IIKRHTGTSEGVTQWMNRLIAFASRGNHVSPTHYIQDDDASKRVMGIL (SEQ ID NO:351); where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0289] In some cases, a TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: ILRRHVGPAEGATQWMNRLIAFASRGNHVSPTHYVPETDASRXVTTIL (SEQ ID NO:350), where X is R or K; where the TP48 T-cell epitope polypeptide has a length of from 40 amino acids to about 48 amino acids (e.g., has a length of 40 amino acids (aa), 41 aa, 42 aa, 43 aa, 44 aa, 45 aa, 46 aa, 47 aa, or 48 aa). In some cases, the TP48 T-cell epitope polypeptide has a length of 48 amino acids. In some cases, the TP is part of a fusion polypeptide, as described in detail below.

[0290] A TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD8 epitopes, e.g., a stretch of about 16 contiguous amino acids having the amino acid sequence: EGAVQWMNRLIAFASR (SEQ ID NO:295), or EGATQWMNRLIAFASR (SEQ ID NO:296), or EGVTQWMNRLIAFASR (SEQ ID NO:297); or a stretch of about 9 contiguous amino acids having the amino acid sequence: WMNRLIAFA (SEQ ID NO:298); or a stretch of about 15 contiguous amino acids having the amino acid sequence: PTHYX1X2X3X4DAX5X6X7VX8(SEQ ID NO:354), where X1is V or I; X2is P, T, A, or Q, X3is E or D; X4is S, T, or D; X5is S or A; X6is A, Q, R, or K; X7is R, K, or X; and X8is T or M.

[0291] A TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD8 epitopes, e.g., a stretch of about 16 contiguous amino acids having the amino acid sequence: EGAVQWMNRLIAFASR (SEQ ID NO:295), or EGATQWMNRLIAFASR (SEQ ID NO:296), or EGVTQWMNRLIAFASR (SEQ ID NO:297); or a stretch of about 9 contiguous amino acids having the amino acid sequence: WMNRLIAFA (SEQ ID NO:298); or a stretch of about 15 contiguous amino acids having the amino acid sequence: PTHYX1X2X3X4DAX5X6X7VX8(SEQ ID NO:355), where X1is V; X2is P, T, or A, X3is E; X4is S or T; X5is S or A; X6is A, Q, or R; X7is R or K; and X8is T.

[0292] A TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure can include one or more CD4 epitopes, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: RHVGPGEGAVQWMNR (SEQ ID NO:299), or RHVGQGEGAVQWMNR (SEQ ID NO:300), or RHIGPGEGAVQWMNR (SEQ ID NO:301), or RHVGPAEGATQWMNR (SEQ ID NO:302), or RHTGTSEGVTQWMNR (SEQ ID NO:303); or a stretch of about 20 contiguous amino acids having the amino acid sequence: HVGPGEGAVQWMNRLIAFAS (SEQ ID NO:304), or HVGQGEGAVQWMNRLIAFAS (SEQ ID NO:305), or HIGPGEGAVQWMNRLIAFAS (SEQ ID NO:306), or HVGPAEGATQWMNRLIAFAS (SEQ ID NO:307), or HTGTSEGVTQWMNRLIAFAS (SEQ ID NO:308); or a stretch of about 20 contiguous amino acids having amino acid sequence: GAVQWMNRLIAFASRGNHVS (SEQ ID NO:309), or GAVQWMNRLIAFASRGNHVA (SEQ ID NO:310), or GATQWMNRLIAFASRGNHVS (SEQ ID NO:311), or GVTQWMNRLIAFASRGNHVS (SEQ ID NO:312).

[0293] In some cases, TP48 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure includes: a) one or more CD8 epitopes, e.g., a stretch of about 16 contiguous amino acids having the amino acid sequence: EGAVQWMNRLIAFASR (SEQ ID NO:295), or EGATQWMNRLIAFASR (SEQ ID NO:296), or EGVTQWMNRLIAFASR (SEQ ID NO:297); or a stretch of about 9 contiguous amino acids having the amino acid sequence: WMNRLIAFA (SEQ ID NO:298); or a stretch of about 15 contiguous amino acids having the amino acid sequence: PTHYX1X2X3X4DAX5X6X7VX8(SEQ ID NO:354), where X1is V or I; X2is P, T, A, or Q, X3is E or D; X4is S, T, or D; X5is S or A; X6is A, Q, R, or K; X7is R, K, or X; and X8is T or M; or a stretch of about 15 contiguous amino acids having the amino acid sequence: PTHYX1X2X3X4DAX5X6X7VX8(SEQ ID NO:355), where X1is V; X2is P, T, or A, X3is E; X4is S or T; X5is S or A; X6is A, Q, or R; X7is R or K; and X8is T; and b) one or more CD4 epitopes, e.g., a stretch of about 15 contiguous amino acids having the amino acid sequence: RHVGPGEGAVQWMNR (SEQ ID NO:299), or RHVGQGEGAVQWMNR (SEQ ID NO:300), or RHIGPGEGAVQWMNR (SEQ ID NO:301), or RHVGPAEGATQWMNR (SEQ ID NO:302), or RHTGTSEGVTQWMNR (SEQ ID NO:303); or a stretch of about 20 contiguous amino acids having the amino acid sequence:HVGPGEGAVQWMNRLIAFAS (SEQ ID NO:304), or HVGQGEGAVQWMNRLIAFAS (SEQ ID NO:305), or HIGPGEGAVQWMNRLIAFAS (SEQ ID NO:306), or HVGPAEGATQWMNRLIAFAS (SEQ ID NO:307), or HTGTSEGVTQWMNRLIAFAS (SEQ ID NO:308); or a stretch of about 20 contiguous amino acids having the amino acid sequence: GAVQWMNRLIAFASRGNHVS (SEQ ID NO:309), or GAVQWMNRLIAFASRGNHVA (SEQ ID NO:310), or GATQWMNRLIAFASRGNHVS (SEQ ID NO:311), or GVTQWMNRLIAFASRGNHVS (SEQ ID NO:312).

[0294] In some cases, a suitable TP is a TP48 polypeptide with from 1 amino acid to 5 amino acids removed from the N-terminus and / or from 1 amino acid to 5 amino acids removed from the C- terminus. Thus, e.g., a suitable TP can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: RHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQIL (SEQ ID NO:356); and has a length of 45 amino acids. As another example, a suitable TP can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVT (SEQ ID NO:357); and has a length of 45 amino acids. TP33 T-cell epitope polypeptides

[0295] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: HSKKKCDELAAKLTGLGLNAVAYYRGLDVSVIP (SEQ ID NO:358); and has a length of from 33 amino acids to about 37 amino acids (e.g., has a length of 33 amino acids (aa), 34 aa, 35 aa, 36 aa, or 37 aa), or has a length of from 30 amino acids to about 33 amino acids (e.g., has a length of 30 amino acids (aa), 31 aa, 32 aa, or 33 aa). In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identityto the following amino acid sequence: HSKKKCDELAAKLTGLGLNAVAYYRGLDVSVIP (SEQ ID NO:358); and has a length of 33 amino acids. In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequence HSKKKCDELAAKLTGLGLNAVAYYRGLDVSVIP (SEQ ID NO:358); and has a length of 33 amino acids. Such TPs are also referred to herein as “TP33” polypeptides. In some cases, a TP33 T-cell epitope polypeptide comprises a CD8 epitope comprising a contiguous stretch of 19 amino acids of one of the following amino acid sequences: i) DELAAKLTGLGLNAVAYYR (SEQ ID NO:359); ii) DELAAKLVALGINAVAYYR (SEQ ID NO:360); iii) DELAAKLSGLGLNAVAYYR (SEQ ID NO:361); iv) DELAAALRGMGLNAVAYYR (SEQ ID NO:362); v) DELAAALRGMGVNAVAYYR (SEQ ID NO:363); vi) DELASKLRGMGLNAVAYYR (SEQ ID NO:364); vii) DELAAKLRGMGLNAVAYYR (SEQ ID NO:365).

[0296] In some cases, a TP33 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: HSKKKCDELAX1X2LX3X4X5GX6NAVAYYRGLDVSX7IP (SEQ ID NO:366), where X1is A or S; X2is K or A; X3is V, S, R, or T; X4is A or G; X5is L or M; X6is I, L, or V; and X7is V or I; and has a length of from 33 amino acids to about 37 amino acids (e.g., has a length of 33 amino acids (aa), 34 aa, 35 aa, 36 aa, or 37 aa), or has a length of from 30 amino acids to about 33 amino acids (e.g., has a length of 30 amino acids (aa), 31 aa, 32 aa, or 33 aa). In some cases, a TP33 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: HSKKKCDELAX1X2LX3X4X5GX6NAVAYYRGLDVSX7IP (SEQ ID NO:366), where X1is A or S; X2is K or A; X3is V, S, R, or T; X4is A or G; X5is L or M; X6is I, L, or V; and X7is V or I; where the TP33 T-cell epitope polypeptide has a length of 33 amino acids.

[0297] In some cases, a TP33 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: HSKKKCDELAAKLX1X2X3GX4NAVAYYRGLDVSVIP (SEQ ID NO:367), where X1is V, S, R, or T; X2is A or G; X3is L or M; and X4is I, L, or V; and has a length of from 33 amino acids to about 37 amino acids (e.g., has a length of 33 amino acids (aa), 34 aa, 35 aa, 36 aa, or 37 aa), or has a length of from 30 amino acids to about 33 amino acids (e.g., has a length of 30 amino acids (aa), 31 aa, 32 aa, or 33 aa). In some cases, a TP33 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: HSKKKCDELAAKLX1X2X3GX4NAVAYYRGLDVSVIP (SEQ ID NO:367), where X1is V, S, R, or T; X2is A or G; X3is L or M; and X4is I, L, or V; where the TP33 T-cell epitope polypeptide has a length of 33 amino acids.

[0298] In some cases, a TP33 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence:HSKKKCDELAAALRGMGX1NAVAYYRGLDVSX2IP (SEQ ID NO:368), where X1is I, L, or V; and X2is V or I; and has a length of from 33 amino acids to about 37 amino acids (e.g., has a length of 33 amino acids (aa), 34 aa, 35 aa, 36 aa, or 37 aa), or has a length of from 30 amino acids to about 33 amino acids (e.g., has a length of 30 amino acids (aa), 31 aa, 32 aa, or 33 aa). In some cases, a TP33 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: HSKKKCDELAAALRGMGX1NAVAYYRGLDVSX2IP (SEQ ID NO:368), where X1is I, L, or V; and X2is V or I; where the TP33 T-cell epitope polypeptide has a length of 33 amino acids.

[0299] In some cases, a TP by an RNA molecule of the present disclosure comprises the amino acid sequence HSKKKCDELAAKLRGMGLNAVAYYRGLDVSVIP (SEQ ID NO:369); and has a length of 33 amino acids. TP42-2 T-cell epitope polypeptides

[0300] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: KGGRHLIFCHSKKKCDELAAKLTGLGLNAVAYYRGLDVSVIP (SEQ ID NO:370); and has a length of from 42 amino acids to about 46 amino acids (e.g., has a length of 42 amino acids (aa), 43 aa, 44 aa, 45 aa, or 46 aa), or has a length of from 38 amino acids to about 42 amino acids (e.g., has a length of 38 amino acids (aa), 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, a T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: KGGRHLIFCHSKKKCDELAAKLTGLGLNAVAYYRGLDVSVIP (SEQ ID NO:370); and has a length of 42 amino acids. In some cases, a TP encoded by an RNA molecule of the present disclosure comprises the amino acid sequence KGGRHLIFCHSKKKCDELAAKLTGLGLNAVAYYRGLDVSVIP (SEQ ID NO:370); and has a length of 42 amino acids. Such TPs are also referred to herein as “TP42-2” polypeptides. In some cases, a TP42-2 T-cell epitope polypeptide comprises a CD8 epitope comprising a contiguous stretch of 19 amino acids of one of the following amino acid sequences: i) DELAAKLTGLGLNAVAYYR (SEQ ID NO:359); ii) DELAAKLVALGINAVAYYR (SEQ ID NO:360); iii) DELAAKLSGLGLNAVAYYR (SEQ ID NO:361); iv) DELAAALRGMGLNAVAYYR (SEQ ID NO:362); v) DELAAALRGMGVNAVAYYR (SEQ ID NO:363); vi)DELASKLRGMGLNAVAYYR (SEQ ID NO:364); vii) DELAAKLRGMGLNAVAYYR (SEQ ID NO:365).

[0301] In some cases, a TP42-2 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KGGRHLIFCHSKKKCDELAX1X2LX3X4X5GX6NAVAYYRGLDVSX7IP (SEQ ID NO:371), where X1is A or S; X2is K or A; X3is V, S, R, or T; X4is A or G; X5is L or M; X6is I, L, or V; and X7is V or I; and has a length of from 42 amino acids to about 46 amino acids (e.g., has a length of 42 amino acids (aa), 43 aa, 44 aa, 45 aa, or 46 aa), or has a length of from 38 amino acids to about 42 amino acids (e.g., has a length of 38 amino acids (aa), 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, a TP42-2 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KGGRHLIFCHSKKKCDELAX1X2LX3X4X5GX6NAVAYYRGLDVSX7IP (SEQ ID NO:371), where X1is A or S; X2is K or A; X3is V, S, R, or T; X4is A or G; X5is L or M; X6is I, L, or V; and X7is V or I; where the TP42-2 T-cell epitope polypeptide has a length of 42 amino acids.

[0302] In some cases, a TP42-2 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KGGRHLIFCHSKKKCDELAAKLX1X2X3GX4NAVAYYRGLDVSVIP (SEQ ID NO:372), where X1is V, S, R, or T; X2is A or G; X3is L or M; and X4is I, L, or V; and has a length of from 42 amino acids to about 46 amino acids (e.g., has a length of 42 amino acids (aa), 43 aa, 44 aa, 45 aa, or 46 aa), or has a length of from 38 amino acids to about 42 amino acids (e.g., has a length of 38 amino acids (aa), 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, a TP42-2 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KGGRHLIFCHSKKKCDELAAKLX1X2X3GX4NAVAYYRGLDVSVIP (SEQ ID NO:372), where X1is V, S, R, or T; X2is A or G; X3is L or M; and X4is I, L, or V; where the TP42-2 T-cell epitope polypeptide has a length of 42 amino acids.

[0303] In some cases, a TP42-2 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: KGGRHLIFCHSKKKCDELAAALRGMGX1NAVAYYRGLDVSX2IP (SEQ ID NO:373), where X1is I, L, or V; and X2is V or I; and has a length of from 42 amino acids to about 46 amino acids (e.g., has a length of 42 amino acids (aa), 43 aa, 44 aa, 45 aa, or 46 aa), or has a length of from 38 amino acids to about 42 amino acids (e.g., has a length of 38 amino acids (aa), 39 aa, 40 aa, 41 aa, or 42 aa). In some cases, a TP42-2 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the following amino acid sequence: HSKKKCDELAAALRGMGX1NAVAYYRGLDVSX1IP (SEQ ID NO:368), where X1is I, L, or V; and X2is V or I; where the TP42-2 T-cell epitope polypeptide has a length of 42 amino acids.

[0304] In some cases, a TP42-2 T-cell epitope polypeptide encoded by an RNA molecule of the present disclosure comprises the amino acid sequence KGGRHLIFCHSKKKCDELAAKLRGMGLNAVAYYRGLDVSVIP (SEQ ID NO:374); and has a length of 42 amino acids. TP65 polypeptides

[0305] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: TPIDTTIMAKNEVFCVDPX6KGGRKPARLIVYPDLGVRVCEKMALYDVVQKLPQAVMGX7SYGF QYS (SEQ ID NO:375), where X6is E, T, or V, and X7is S, A, or P; and has a length of from 65 amino acids to 70 amino acids (e.g., 65, 66, 67, 68, 69, or 70 amino acids). In some cases, the TP has a length of 65 amino acids. In some cases, a TP65 polypeptide has the following amino acid sequence: TPIDTTIMAKNEVFCVDPEKGGRKPARLIVYPDLGVRVCEKMALYDVVQKLPQAVMGSSYGFQ YS (SEQ ID NO:376) and has a length of 65 amino acids. In some cases, a TP65 polypeptide has the following amino acid sequence: TPIDTTIMAKNEVFCVDPTKGGRKPARLIVYPDLGVRVCEKMALYDVVQKLPQAVMGASYGFQ YS (SEQ ID NO:377) and has a length of 65 amino acids. In some cases, a TP65 polypeptide has the following amino acid sequence: TPIDTTIMAKNEVFCVDPVKGGRKPARLIVYPDLGVRVCEKMALYDVVQKLPQAVMGPSYGFQ YS (SEQ ID NO:378). TP240 polypeptides

[0306] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: SYQVGYLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRT VTTGAPITYSTYGKFLADGGCSGGAYDIIICDECHSX1DATTILGIGTVLDQAETAGARLVVLATA TPPGSVTVPHPNIEEVALGX2EGEIPFYGKAIPLX3X4IKGGRHLIFCHSKKKCDELAAKLRGMGLN AVAYYRGLDVSVIPTX5GDVVVVATDALMTGYTGDFDSVIDCNVAVTQT (SEQ ID NO:379), where X1is T, V, or Q, X2is T, N, Q, H, or S, X3is E, S, or A, X4is V, T, Y, F, or L, and X5is S, Q, orT; and has a length of from 240 amino acids to 245 amino acids (e.g., 240, 241, 242, 243, 244, or 245 amino acids). In some cases, the TP has a length of 240 amino acids.

[0307] In some cases, a TP240 polypeptide has the following amino acid sequence: SYQVGYLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRT VTTGAPITYSTYGKFLADGGCSGGAYDIIICDECHSTDATTILGIGTVLDQAETAGARLVVLATA TPPGSVTVPHPNIEEVALGTEGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLRGMGLNA VAYYRGLDVSVIPTSGDVVVVATDALMTGYTGDFDSVIDCNVAVTQT (SEQ ID NO:380), and has a length of 240 amino acids. In some cases, a TP240 polypeptide has the following amino acid sequence: SYQVGYLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRT VTTGAPITYSTYGKFLADGGCSGGAYDIIICDECHSVDATTILGIGTVLDQAETAGARLVVLATA TPPGSVTVPHPNIEEVALGNEGEIPFYGKAIPLSTIKGGRHLIFCHSKKKCDELAAKLRGMGLNA VAYYRGLDVSVIPTQGDVVVVATDALMTGYTGDFDSVIDCNVAVTQT (SEQ ID NO:381), and has a length of 240 amino acids. TP29 T-cell epitope polypeptides

[0308] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP29 amino acid sequence depicted in FIG.4; and has a length of about 29 amino acids. For example, in some cases, a TP29 polypeptide comprises the amino acid sequence: AIPLX(EA)X(VTYFL)IKGGRHLIFCHSKKKCDELAAKL (SEQ ID NO:52), where the first “X” is Glu or Ala, and the second “X” is Val, Thr, Tyr, Phe, or Leu; and where the TP29 polypeptide has a length of 29 amino acids. TP50-NS2 T-cell epitope polypeptides

[0309] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP50-NS2 amino acid sequence depicted in FIG.4; and has a length of about 50 amino acids. For example, in some cases, a TP50-NS2 polypeptide comprises the amino acid sequence: LGRWTGTYX(VI)YDHLTPLX(RS)DWAAAGLRDLAVAVEPVVFSPMEX(TK)KVITWGADT(SEQ ID NO:53), where the first “X” is Val or Ile, the second “X” is Arg or Ser, and the third “X” is Thr or Lys; and where the TP50-NS2 polypeptide has a length of 50 amino acids. TP52 T-cell epitope polypeptides

[0310] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP52 amino acid sequence depicted in FIG.4; and has a length of about 52 amino acids. For example, in some cases, a TP52 polypeptide comprises the amino acid sequence: AIPLX(EA)X(VTYFL)IKGGRHLIFCHSKKKCDELAAKLRGMGLNAVAYYRGLDVSVIPT X(SQ) (SEQ ID NO:54), where the first “X” is Glu or Ala, the second “X” is Val, Thr, Tyr, Phe, or Leu, and the third “X” is Ser or Gln; and where the TP52 polypeptide has a length of 52 amino acids. TP70 T-cell epitope polypeptides

[0311] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP70 amino acid sequence depicted in FIG.4; and has a length of about 70 amino acids. For example, in some cases, a TP70 polypeptide comprises the TP70 amino acid sequence depicted in FIG.4, where X is Ser or Gln; and where the TP70 polypeptide has a length of 70 amino acids. TP100 T-cell epitope polypeptides

[0312] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP100 amino acid sequence depicted in FIG.4; and has a length of about 100 amino acids. For example, in some cases, a TP100 polypeptide comprises the TP100 amino acid sequence depicted in FIG.4, where the first “X” is Thr, Asn, Gln, His, or Ser, where the second “X” is Glue or Ala, the third “X” is Val, Thr, Tyr, Phe, or Leu, and the fourth “X” is Ser or Gln; and where the TP100 polypeptide has a length of 100 amino acids. TP171 T-cell epitope polypeptides

[0313] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP171 amino acid sequence depicted in FIG.4; and has a length of about 171 amino acids. TP228 T-cell epitope polypeptides

[0314] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP228 amino acid sequence depicted in FIG.4; and has a length of about 228 amino acids.

[0315] A TP228 polypeptide can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following TP228 amino acid sequence: LHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRTVTTGAP ITYSTYGKFLADGGCSGGAYDIIICDECHSX1DATTILGIGTVLDQAETAGARLVVLATATPPGSV TVPHPNIEEVALGX2EGEIPFYGKAIPLX3X4IKGGRHLIFCHSKKKCDELAAKLRGMGLNAVAYY RGLDVSVIPTX5GDVVVVATDALMTGYTGDFDSVIDCN (SEQ ID NO:58), where X1is Thr or Val; X2is Thr, Asn, Gln, His, or Ser; X3is Glu or Ala; X4is Val, Thr, Tyr, Phe, or Leu; and X5is Ser or Gln; and can have a length of 228 amino acids.

[0316] For example, a TP228 polypeptide can comprise an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: LHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRTITTGSPI TYSTYGKFLADGGCSGGAYDIIICDECHSTDATSILGIGTVLDQAETAGARLVVLATATPPGSVT VPHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGL DVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCN (SEQ ID NO:382); and can have a length of from 215 amino acids (aa) to 235 aa (e.g., 215 aa, 216 aa, 21711, 218 aa, 219 aa, 220 aa, 221 aa, 222 aa, 223 aa, 224 aa, 225 aa, 226 aa, 227 aa, 228 aa, 229 aa, 230 aa, 231 aa, 232 aa, 233 aa, 234 aa, or 235 aa). In some cases, a TP228 polypeptide comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence: LHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRTITTGSPI TYSTYGKFLADGGCSGGAYDIIICDECHSTDATSILGIGTVLDQAETAGARLVVLATATPPGSVT VPHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGL DVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCN (SEQ ID NO:382); and has a length of 228 amino acids. TP553 T-cell epitope polypeptides

[0317] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP553 amino acid sequence depicted in FIG.4; and has a length of about 553 amino acids. TP778 T-cell epitope polypeptides

[0318] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP778 amino acid sequence depicted in FIG.4; and has a length of about 778 amino acids. TP1987 T-cell epitope polypeptides

[0319] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the TP1987 amino acid sequence depicted in FIG.4; and has a length of about 1987 amino acids. TP465 polypeptides

[0320] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to any one of the TP465polypeptide amino acid sequences depicted in FIG.32B, FIG.33B, and FIG.34B; and has a length of from 460 amino acids to 470 amino acids, e.g., has a length of 465 amino acids.

[0321] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0322] SFQVAHLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAH GVDPNIRTGVRTITTGAPITYSTYGKFLADGGCSGGAYDIIICDECHSTDATSILGIGTVLDQ AETAGARLVVLATATPPGSVTVPHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKK KCDELAAKLVALGINAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNTCVT QTVDFSLDPTFTIETTTLPQDAVSRTQRRGRTGRGKPGIYRFVAPGERPSGMFDSSVLCECY DAGCAWYELTPAETTVRLRAYMNTPGLPVCQDHLEFWEGVFTGLTHIDAHFLSQTKQSGE NFPYLVAYQATVCARAQAPPPSWDQMWKCLIRLKPTLHGPTPLLYRLGAVQNEVTLTHPVT KYIMTCMSADLEVVTSTWVLVGGVLAALAAYCLSTGCVVIVGRI (SEQ ID NO:383); and has a length of about 465 amino acids. TP156 polypeptides

[0323] In some cases, a TP encoded by an RNA molecule of the present disclosure comprises an amino acid sequence having at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0324] KFPGGGQIVGGVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRPEGRTW AQPGYPWPLYGNEGCGWAGWLLSPRGSRPSWGPTDPRRRSRNLGKVIDTLTCGFADLMGYIPL VGAPLGGAARALAHGVRVLEDGVNYATGNLPGCSFSIFL (SEQ ID NO:384); and has a length of about 165 amino acids. Multiple T-cell epitope polypeptides in a single polypeptide chain (fusion polypeptides)

[0325] In some cases, a heterologous polypeptide encoded in an RNA molecule of the present disclosure is a fusion polypeptide comprising 2, 3, 4, 5, 6, 7, 8, 9, or 10 of the aforementioned TPs in a single polypeptide chain. In other words, the heterologous polypeptide is in some cases a fusion polypeptide comprising 2, 3, 4, 5, 6, 7, 8, 9, or 10 of the TPs in a single polypeptide chain, where such TPs may be referred to as a “TP fusion polypeptide.”

[0326] For example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP35-NS3 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and having a length of 35 amino acids); and ii) a TP50C polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and having a length of 50 amino acids. Thus, e.g., in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising a polypeptide having at least 20% (at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or 100%) amino acid identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKGVYLLPRRGPRLGVRATRKTSERSQPR GRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:385) and having a length of 85 amino acids.

[0327] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP50C polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and having a length of 50 amino acids; and ii) a TP35-NS3 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and having a length of 35 amino acids). Thus, e.g., in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding s: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising a polypeptide having at least 20% (at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or 100%) amino acid identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYPKSTKVPAAYAAQG YKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:386) and having a length of 85 amino acids.

[0328] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP35-NS3 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and having a length of 35 amino acids); ii) a TP50C polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and having a length of 50 amino acids; and iii) a TP23 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and having a length of 23 amino acids). Thus, e.g., in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide having at least 20% (at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or 100%) amino acid identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKGVYLLPRRGPRLGVRATRKTSERSQPR GRRQPIPKARRSEGRSWAQPGYPDVVVVATDALMTGFTGDFDSVID (SEQ ID NO:387) and having a length of 108 amino acids.

[0329] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP35-NS3 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and having a length of 35 amino acids); ii) a TP50C polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and having a length of 50 amino acids; iii) a TP23 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and having a length of 23 amino acids); and iv) a TP27 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272); and having a length of 27 amino acids). Thus, e.g., in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising an amino acid sequence having at least 20% (at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or 100%) amino acid identity to:KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKGVYLLPRRGPRLGVRATRKTSERSQPR GRRQPIPKARRSEGRSWAQPGYPDVVVVATDALMTGFTGDFDSVIDLEQIKGGRHLIFCHSKKK CDELAAKLT SEQ ID NO:388) and having a length of 135 amino acids.

[0330] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP35-NS3 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and having a length of 35 amino acids); ii) a TP50C polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and having a length of 50 amino acids; iii) a TP23 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and having a length of 23 amino acids); iv) a TP27 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272); and having a length of 27 amino acids); and v) a TP35-NS4 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287); and having a length of 35 amino acids. Thus, e.g., in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide having at least 20% (at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or 100%) amino acid identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKGVYLLPRRGPRLGVRATRKTSERSQPR GRRQPIPKARRSEGRSWAQPGYPDVVVVATDALMTGFTGDFDSVIDLEQIKGGRHLIFCHSKKK CDELAAKLTILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:389) and having a length of 170 amino acids.

[0331] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP35-NS3 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and having a length of 35 amino acids); ii) a TP50C polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP(SEQ ID NO:101); and having a length of 50 amino acids; iii) a TP42 polypeptide as described above (e.g., a polypeptide having following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG (SEQ ID NO:313); and having a length of 42 amino acids); and iv) a TP48 polypeptide as described above (e.g., a polypeptide having following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); and having a length of 48 amino acids).

[0332] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; and b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP35-NS3 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and having a length of 35 amino acids); ii) a TP50C polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and having a length of 50 amino acids; iii) a TP42 polypeptide as described above (e.g., a polypeptide having following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG (SEQ ID NO:303); and having a length of 42 amino acids); iv) a TP48 polypeptide as described above (e.g., a polypeptide having following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); and having a length of 48 amino acids); and v) a TP42-2 polypeptide, as described above (e.g., a polypeptide having the following amino acid sequence: KGGRHLIFCHSKKKCDELAAKLTGLGLNAVAYYRGLDVSVIP (SEQ ID NO:370); and having a length of 42 amino acids.

[0333] A TP fusion polypeptide encoded by an RNA molecule of the present disclosure can comprise: i) a TP50-C polypeptide having the amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); ii) a TP27 polypeptide having the amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLR (SEQ ID NO:103); iii) a TP48 polypeptide having the amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); and iv) a TP35-NS3 polypeptide having the amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102), where the TP50C-, TP27, TP48, and TP35-NS3 polypeptides are in any order in the TP fusion polypeptide.

[0334] As an example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; and b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP50-C polypeptide having the amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); ii) a TP35-NS3 polypeptide having the amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); iii) a TP27 polypeptide having the amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLR (SEQ ID NO:103); and iv) a TP48 polypeptide having the amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104).

[0335] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; and b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP48 polypeptide having the amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); ii) a TP50-C polypeptide having the amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); iii) a TP35-NS3 polypeptide having the amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and iv) a TP27 polypeptide having the amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLR (SEQ ID NO:103).

[0336] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; and b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP27 polypeptide having the amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLR (SEQ ID NO:103); ii) a TP48 polypeptide having the amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); iii) a TP50-C polypeptide having the amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and iv) a TP35-NS3 polypeptide having the amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102).

[0337] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; and b) a TPfusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP35-NS3 polypeptide having the amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); ii) a TP27 polypeptide having the amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLR (SEQ ID NO:103); iii) a TP48 polypeptide having the amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); and iv) a TP50-C polypeptide having the amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101).

[0338] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; and b) a TP fusion polypeptide comprising, in order from N-terminus to C-terminus: i) a TP50-C polypeptide having the amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); ii) a TP27 polypeptide having the amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLR (SEQ ID NO:103); iii) a TP48 polypeptide having the amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDAAARVTQIL (SEQ ID NO:104); and iv) a TP35-NS3 polypeptide having the amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:104).

[0339] As another example, in some cases, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; and b) a TP fusion polypeptide comprising: i) a TP240 polypeptide; ii) a TP48 polypeptide; and iii) a TP65 polypeptide; iv) a TP50C polypeptide. The TP240 polypeptide, the TP48 polypeptide, the TP65 polypeptide, and the TP50C polypeptide can be in the TP fusion polypeptide in any order. For example, a composition of the present disclosure comprises one or more RNA molecules comprising a nucleotide sequence encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; and b) a TP fusion polypeptide comprising, in order from N-terminus to C- terminus: i) a TP240 polypeptide; ii) a TP48 polypeptide; and iii) a TP65 polypeptide; iv) a TP50C polypeptide. As an example, the TP fusion polypeptide can comprise an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity, to the amino acid sequence depicted in FIG.29, where X1is T, V, or Q, X2is T, N, Q, H, or S, X3is E, S, or A, X4is V, T, Y, F, or L, X5is S, Q, or T, X6is E, T, or V, and X7is S, A, or P; where the TP fusion polypeptide has a length of from 410 amino acids to 415 amino acids (e.g., 410, 411, 412, 413, 414, or 415 amino acids). In some cases, the TP fusion polypeptide has a length of 410 amino acids.Combinations

[0340] As noted above, an RNA molecule of the present disclosure can comprise (in addition to a nucleotide sequence(s) encoding an HCV E1 and / or E2 polypeptide) nucleotide sequences encoding 2, 3, 4, 5, 6, 7, 8, 9, or 10 different T-cell epitope polypeptides as the heterologous polypeptide.

[0341] Combination #1. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first T-cell epitope polypeptide comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101). The first TP can have a length of 35 amino acids; and the second TP can have a length of 50 amino acids. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; and c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids.

[0342] Combination #2. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); c) a second T-cell epitope polypeptide comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and d) a pharmaceutically acceptable excipient. The first TP can have a length of 35 amino acids; and the second T-cell epitope polypeptide can have a length of 23 amino acids. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; and c) a second TP comprising the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids.

[0343] Combination #3. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272). The first TP can have a length of 35 amino acids; and the second TP can have a length of 27 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids.

[0344] Combination #4. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first TP can have a length of 35 amino acids; and the second TP can have a length of 35 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; and c) a second TP comprising the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0345] Combination #5. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256). The first TP can have a length of 50 amino acids; and the second TP can have a length of 23 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising thefollowing amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; and c) a second T-cell epitope polypeptide comprising the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids;

[0346] Combination #6. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and c) a second T-cell epitope polypeptide comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272). The first T-cell epitope polypeptide can have a length of 50 amino acids; and the second T-cell epitope polypeptide can have a length of 27 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first T-cell epitope polypeptide comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; and c) a second T-cell epitope polypeptide comprising the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids.

[0347] Combination #7. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first T-cell epitope polypeptide can have a length of 50 amino acids; and the second TP can have a length of 35 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; and c) a second TP comprising the following amino acid sequence:ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0348] Combination #8. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272). The first TP can have a length of 23 amino acids; and the second T-cell epitope polypeptide can have a length of 27 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids; and c) a second TP comprising the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids.

[0349] Combination #9. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and c) a second T-cell epitope polypeptide comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first TP can have a length of 23 amino acids; and the second TP can have a length of 35 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids; and c) a second TP comprising the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0350] Combination #10. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). Thefirst T-cell epitope polypeptide can have a length of 27 amino acids; and the second T-cell epitope polypeptide can have a length of 35 amino acids. As an example, an immunogenic composition of the present disclosure can comprise: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids; and c) a second TP comprising the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0351] Combination #11. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256). The first TP can have a length of 35 amino acids; the second TP can have a length of 50 amino acids; and the third TP can have a length of 23 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids.

[0352] Combination #12. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272). The first TP can have a length of 35 amino acids; the second TP can have a length of 50 amino acids; and the third TP canhave a length of 27 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids.

[0353] Combination #13. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first TP can have a length of 35 amino acids; the second TP can have a length of 50 amino acids; and the third TP can have a length of 35 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0354] Combination #14. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to:LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272). The first TP can have a length of 50 amino acids; the second TP can have a length of 23 amino acids; and the third TP can have a length of 27 amino acids. As an example an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; c) a second TP comprising the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids; and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids.

[0355] Combination #15. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first TP can have a length of 50 amino acids; the second TP can have a length of 23 amino acids; and the third TP can have a length of 35 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; c) a second TP comprising the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids; and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0356] Combination #16. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272); and d) a third TP comprising an aminoacid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first TP can have a length of 23 amino acids; the second TP can have a length of 27 amino acids; and the third TP can have a length of 35 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids; c) a second TP comprising the following amino acid sequence: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids; and d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0357] Combination #17. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and e) a fourth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272). The first TP can have a length of 35 amino acids; the second TP can have a length of 50 amino acids; the third TP can have a length of 23 amino acids; and the fourth TP can have a length of 27 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids; and e) a fourth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids.

[0358] Combination #18. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); and e) a fourth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first TP can have a length of 35 amino acids; the second TP can have a length of 50 amino acids; the third TP can have a length of 23 amino acids; and the fourth TP can have a length of 35 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids; and e) a fourth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0359] Combination #19. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272); and e) a fourth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first TP can have a length of 35 amino acids; the second TP can have a length of 50 amino acids; the third TP can have a length of 27 amino acids; and the fourth TP can have a length of 35 amino acids. As an example, anRNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having a length of 27 amino acids; and e) a fourth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0360] Combination #20. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101); d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256); e) a fourth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272); and f) a fifth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287). The first TP can have a length of 35 amino acids; the second TP can have a length of 50 amino acids; the third TP can have a length of 23 amino acids; the fourth TP can have a length of 27 amino acids; and the fifth TP can have a length of 35 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101) and having a length of 50 amino acids; d) a third TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: DVVVVATDALMTGFTGDFDSVID (SEQ ID NO:256) and having a length of 23 amino acids; e) a fourth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LEQIKGGRHLIFCHSKKKCDELAAKLT (SEQ ID NO:272) and having alength of 27 amino acids; and f) a fifth TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYV (SEQ ID NO:287) and having a length of 35 amino acids.

[0361] Combination #21. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG(K)n (SEQ ID NO:390), where n is an integer from 2 to 10 (e.g., where n is 2, 3, 4, 5, 6, 7, 8, 9, or 10). The first TP can have a length of 35 amino acids; and the second TP can have a length of from 44 amino acids to 52 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; and c) a second TP comprising the following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTGKKK (SEQ ID NO:391) and has a length of 45 amino acids.

[0362] Combination #22. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNV(K)n (SEQ ID NO:392), where n is an integer from 2 to 10 (e.g., where n is 2, 3, 4, 5, 6, 7, 8, 9, or 10). The first TP can have a length of 35 amino acids; and the second TP can have a length of from 47 amino acids to 57 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; and c) a second TP comprising the following amino acid sequence: LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNVKKK (SEQ ID NO:334) and having a length of 48 amino acids.

[0363] Composition #23. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQIL(Lys)n (SEQ ID NO:393), where n is an integer from 2 to 10 (e.g., where n is 2, 3, 4, 5, 6, 7, 8, 9, or 10). The first TP can have a length of 35 amino acids; and the second TP can have a length of from 50 amino acids to 58 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: KSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK (SEQ ID NO:102) and having a length of 35 amino acids; c) a second TP comprising the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQILKKK (SEQ ID NO:394) and having a length of 51 amino acids.

[0364] Combination #24. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG(K)n (SEQ ID NO:390), where n is an integer from 2 to 10 (e.g., where n is 2, 3, 4, 5, 6, 7, 8, 9, or 10); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNV(K)n (SEQ ID NO:392), where n is an integer from 2 to 10 (e.g., where n is 2, 3, 4, 5, 6, 7, 8, 9, or 10). The first TP can have a length of from 44 amino acids to 52 amino acids; and the second TP can have a length of from 47 amino acids to 57 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTGKKK (SEQ ID NO:391) and having a length of 45 amino acids; and c) a second TP comprising the following amino acid sequence: LNAVAYYRGLDVSVIPTSGDVVVVATDALMTGFTGDFDSVIDCNVKKK (SEQ ID NO:394) and having a length of 48 amino acids.

[0365] Combination #25. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCVE2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG(K)n (SEQ ID NO:390), where n is an integer from 2 to 10 (e.g., where n is 2, 3, 4, 5, 6, 7, 8, 9, or 10); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQIL (SEQ ID NO:338). The first TP can have a length of from 44 amino acids to 52 amino acids; and the second TP can have a length of 48 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTGKKK (SEQ ID NO:391) and having a length of 45 amino acids; and c) a second TP comprising the following amino acid sequence: ILRRHVGPGEGAVQWMNRLIAFASRGNHVSPTHYVPESDASARVTQIL(Lys)n (SEQ ID NO:393), where n is an integer from 2 to 10 (e.g., where n is 2, 3, 4, 5, 6, 7, 8, 9, or 10).

[0366] Combination #26. As one example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTG(K)n (SEQ ID NO:390), where n is an integer from 2 to 10 (e.g., where n is 2, 3, 4, 5, 6, 7, 8, 9, or 10); and c) a second TP comprising an amino acid sequence having at least 20% amino acid sequence identity to: GVYLLPRRGPRLGVRATRKTSERSQPRGRRQPIPKARRSEGRSWAQPGYP (SEQ ID NO:101). The first TP can have a length of from 44 amino acids to 52 amino acids; and the second TP can have a length of 50 amino acids. As an example, an RNA molecule of the present disclosure comprises nucleotide sequences encoding: a) i) an HCV E1 / E2 heterodimeric polypeptide; ii) an HCV E2 polypeptide; or iii) an HCV E1 polypeptide; b) a first TP comprising the following amino acid sequence: GTEGEIPFYGKAIPLEQIKGGRHLIFCHSKKKCDELAAKLTGKKK (SEQ ID NO:391) and having a length of 45 amino acids; and c) a second TP comprising the following amino acid sequence: GVYLLPRRGPRLG...

Claims

CLAIMS What is claimed is:

1. A composition comprising: a) one or more RNA molecules, wherein the one or more RNA molecules comprises: i) at least one 5’ cap structure and / or a 5’ untranslated region (5’ UTR) and / or a 5’ internal ribosome entry site (IRES); ii) a polypeptide-encoding region comprising one or more nucleotide sequences encoding a hepatitis C virus (HCV) E1 polypeptide and / or an HCV E2 polypeptide; and b) one or more of: i) a lipid; ii) a polymer; iii) a peptide; iv) a viral-like particle; and v) a cationic nanoemulsion.

2. The composition of claim 1, wherein the one or more RNA molecules comprises a 5’ cap structure and a 5’ UTR, wherein the 5’UTR is 3’ of the 5’ cap structure.

3. The composition of claim 1 or claim 2, wherein the one or more RNA molecules comprises a 3’ UTR and / or a 3’ tailing sequence.

4. The composition of claim 3, wherein the 3’ tailing sequence is a poly(adenosine) (poly- A) sequence.

5. The composition of any one of claims 1-4, wherein the one or more RNA molecules include at least one 5’ cap structure, and wherein the at least one 5’ cap structure is selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2'fluoro-guanosine, 7-deaza- guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

6. The composition of any one of claims 1-5, wherein the one or more RNA molecules comprises one or more of: a nucleoside base modification, a sugar modification, and a backbone modification.

7. The composition any one of claims 1-6, wherein at least one of the one or more RNA molecules comprises a nucleotide of Formula I:wherein: U is O, S, N(RU)nu, or C(RU)nu,where nu is an integer nu is an integer from 0 to 2 and each RUis, independently, H, halo, or optionally substituted alkyl; is a single or double bond; is a single bond or absent; each of R1', R2', R1'', R2'', R3, R4, and R5is, independently, H, halo, hydroxy, thiol, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted aminoalkoxy, optionally substituted alkoxyalkoxy, optionally substituted hydroxyalkoxy, optionally substituted amino, azido, optionally substituted aryl, optionally substituted aminoalkyl, or absent; wherein the combination of R3with one or more of R1', R1'', R2', R2'', or R5can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl; wherein the combination of R5with one or more of R1', R1'', R2', or R2''can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl; and wherein the combination of R4and one or more of R1', R1'', R2', R2'', R3, or R5can join together to form optionally substituted alkylene or optionally substituted heteroalkylene and, taken together with the carbons to which they are attached, provide an optionally substituted heterocyclyl; each of Y1, Y2, and Y3, is, independently, O, S, -NRN1-, optionally substituted alkylene, or optionally substituted heteroalkylene, wherein RN1is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, or absent; each Y4is, independently, H, hydroxy, thiol, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted alkenyloxy, optionally substituted alkynyloxy, optionally substituted thioalkoxy, optionally substituted alkoxyalkoxy, or optionally substituted amino; each Y5is, independently, O, S, optionally substituted alkylene, or optionally substituted heteroalkylene;n is an integer from 1 to 100,000; and B is a nucleobase, wherein the combination of B and R1', the combination of B and R2', the combination of B and R1'', or the combination of B and R2'' can, taken together with the carbons to which they are attached, optionally form a bicyclic group or wherein the combination of B, R1'', and R3or the combination of B, R2'', and R3can optionally form a tricyclic or tetracyclic group.

8. The composition of any one of claims 1-7, wherein the composition comprises a lipid, and wherein the lipid is selected from DLin-DMA, DLin-K-DMA, DLin-KC2-DMA, 98N12-5, C12-200, DLin-MC3-DMA, DODMA, DSDMA, DLenDMA, reLNPs, PLGA, a PEGylated lipid, and mixtures of two or more of the foregoing.

9. The composition of any one of claims 1-7, wherein the composition comprises a polymer.

10. The composition of claim 9, herein the polymer is selected from polyethylene glycol, polyglycolide, polyvinyl alcohol, polyvinyl pyrrolidone, polylactide, poly(lactide-co-glycolide) (PLGA), polycaprolactone, polysorbate, polyethylene oxide, polypropylene oxide, poly(ethylene oxide-co- propylene oxide), poloxamer, poloxamine, poly(oxyethylated) glycerol, poly(oxyethylated) sorbitol, poly(oxyethylated) glucose, polyethyleneimine, polyamidoamine (PAMAM) dendrimer, and block copolymer poly(ethylene glycol)-block-poly(lactic-co-glycolic acid) (PEG-b-PLGA).

11. The composition of any one of claims 1-10, wherein the HCV E1 polypeptide is a soluble HCV E1 polypeptide.

12. The composition of any one of claims 1-10, wherein the HCV E2 polypeptide is a soluble HCV E2 polypeptide.

13. The composition of any one of claims 1-12, wherein the nucleotide sequence encodes an HCV E1 polypeptide and an HCV E2 polypeptide.

14. The composition of claim 13, wherein: a) the HCV E1 polypeptide is a soluble HCV E1 polypeptide; b) the HCV E2 polypeptide is a soluble HCV E2 polypeptide; or c) the HCV E1 polypeptide is a soluble HCV E1 polypeptide and the HCV E2 polypeptide is a soluble HCV E2 polypeptide.

15. The composition of any one of claims 1-14, wherein the polypeptide-encoding region comprises a nucleotide sequence encoding an HCV Core polypeptide.

16. The composition of claim 15, wherein the polypeptide-encoding region comprises a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein.

17. The composition of any one of claims 1-16, wherein the polypeptide-encoding region comprises: a) one or more nucleotide sequences encoding one or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or b) a nucleotide sequence encoding a fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

18. The composition of claim 17, wherein the RNA comprises: a) an internal ribosome entry site (IRES) between i) the one or more nucleotide sequences encoding the HCV E1 and / or E2 polypeptides; and ii) the one or more nucleotide sequences encoding the one or more heterologous polypeptides or the nucleotide sequence encoding the fusion polypeptide; or b) a nucleotide sequence encoding a self-cleaving peptide between i) the one or more nucleotide sequences encoding the HCV E1 and / or E2 polypeptides; and ii) the one or more nucleotide sequences encoding the one or more heterologous polypeptides or the nucleotide sequence encoding the fusion polypeptide.

19. The composition of claim 17 or claim 18, wherein the one or more heterologous polypeptides is selected from: i) a TP35-NS3 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), wherein X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R, wherein the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; ii) a TP50C T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGYP (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Qor K, wherein the TP50C T-cell epitope polypeptide has a length of from 40 amino acids to 50 amino acids; iii) a TP23 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: DVVVX1X2TDALMTGX3TGDFDSVID (SEQ ID NO:257), wherein X1is V or C; X2is A or S; and X3is F or Y, wherein the TP23 T-cell epitope polypeptide has a length of from 18 amino acids to 23 amino acids; iv) a TP27 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S, wherein the TP27 T-cell epitope polypeptide has a length of from 22 amino acids to 27 amino acids; v) a TP35-NS4 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10(SEQ ID NO:288), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; and X10is V or I, wherein the TP35-NS4 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; vi) a TP42 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3GEIPFYGX4AIPX5X6X7X8KGGRHLIFCHSKKKCDEX9AX10X11LX12X13(K)n (SEQ ID NO:314), where X1is G, P, or S; X2is T, N, Q, H, or S; X3is E, T, or D; X4is K or R; X5is L or I; X6is E, A, S, or Q; X7is Q, T, Y, F, or L; X8is I or L; X9is L or I; X10is A, K, or S; X11is K, Q, or A; X12is T, R, or S; and X13is G or S, wherein n is an integer from 2 to 10, and wherein the TP42 T-cell epitope polypeptide has a length of from 34 amino acids to 52 amino acids; vii) a TP45 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2AVAX3YRGX4DVX5X6IPX7X8GDVVVX9X10TDALMTGX11TGDFDSVIDX12X13X14(K)n (SEQ ID NO:332), wherein X1is L or V; X2is N or T; X3is Y or F; X4is L or V; X5is S or A; X6is V or I; X7is T or A; X8is S, Q, or T; X9is V or C; X10is A or S; X11is F or Y; X12is C or K; X13is N or K; and X14is V or K, wherein n is an integer from 2 to 10, and wherein the TP45 T-cell epitope polypeptide has a length of from 36 amino acids to 55 amino acids; and viii) a TP48 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, orT; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V, wherein n is an integer from 2 to 10, and wherein the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acids; ix) a TP33 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: HSKKKCDELAX1X2LX3X4X5GX6NAVAYYRGLDVSX7IP (SEQ ID NO:366), where X1is A or S; X2is K or A; X3is V, S, R, or T; X4is A or G; X5is L or M; X6is I, L, or V; and X7is V or I; wherein the TP33 T-cell epitope polypeptide has a length of from 30 amino acids to 36 amino acids; x) a TP42-2 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KGGRHLIFCHSKKKCDELAX1X2LX3X4X5GX6NAVAYYRGLDVSX7IP (SEQ ID NO:371), where X1is A or S; X2is K or A; X3is V, S, R, or T; X4is A or G; X5is L or M; X6is I, L, or V; and X7is V or I; wherein the TP42-2 T-cell epitope polypeptide has a length of from 38 amino acids to 46 amino acids; xi) a TP240 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: SYQVGYLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRT VTTGAPITYSTYGKFLADGGCSGGAYDIIICDECHSX1DATTILGIGTVLDQAETAGARLVVLATA TPPGSVTVPHPNIEEVALGX2EGEIPFYGKAIPLX3X4IKGGRHLIFCHSKKKCDELAAKLRGMGLN AVAYYRGLDVSVIPTX5GDVVVVATDALMTGYTGDFDSVIDCNVAVTQT (SEQ ID NO:379), where X1is T, V, or Q, X2is T, N, Q, H, or S, X3is E, S, or A, X4is V, T, Y, F, or L, and X5is S, Q, or T; wherein the TP240 T-cell epitope polypeptide has a length of from 240 amino acids to 245 amino acids; xii) a TP65 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: TPIDTTIMAKNEVFCVDPX6KGGRKPARLIVYPDLGVRVCEKMALYDVVQKLPQAVMGX7SYGF QYS (SEQ ID NO:375), where X6is E, T, or V, and X7is S, A, or P; and wherein the TP65 T-cell epitope polypeptide has a length of from 65 amino acids to 70 amino acids; xiii) a TP156 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP156 amino acid sequence depicted in FIG.32C, where the TP156 T-cell epitope polypeptide has a length of from 150 amino acids to 160 amino acids; and xiv) a TP465 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 amino acid sequence depicted in any one of FIG.32B, 33B, and 34B, where the TP465 T-cell epitope polypeptide has a length of from 460 amino acids to 470 amino acids.

20. The composition of any one of claims 17-19, wherein the polypeptide-encoding region comprises one or more nucleotide sequences encoding two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

21. The composition of claim 20, wherein the mRNA comprises: a) a first internal ribosome entry site (IRES) between i) the one or more nucleotide sequences encoding the HCV E1 and / or E2 polypeptides; and ii) the nucleotide sequence encoding the first of the two or more heterologous polypeptides; and b) a second IRES between i) the nucleotide sequence encoding the first of the two or more heterologous polypeptides; and ii) the nucleotide sequence encoding the second of the two or more heterologous polypeptides, wherein the first IRES and the second IRES are the same or different.

22. The composition of claim 21, wherein the mRNA comprises an IRES between any two consecutive nucleotide sequences encoding heterologous polypeptides.

23. The composition of claim 17 or claim 18, wherein fusion polypeptide comprises from 2 to 10 polypeptides selected from the group consisting of: i) a TP35-NS3 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), wherein X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R, wherein the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; and ii) a TP50C T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGY P (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K, wherein the TP50C T-cell epitope polypeptide has a length of from 40 amino acids to 50 amino acids; iii) a TP23 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: DVVVX1X2TDALMTGX3TGDFDSVID (SEQ ID NO:257), whereinX1is V or C; X2is A or S; and X3is F or Y, wherein the TP23 T-cell epitope polypeptide has a length of from 18 amino acids to 23 amino acids; iv) a TP27 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S, wherein the TP27 T-cell epitope polypeptide has a length of from 22 amino acids to 27 amino acids; v) a TP35-NS4 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10(SEQ ID NO:288), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; and X10is V or I, wherein the TP35-NS4 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; vi) a TP42 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3GEIPFYGX4AIPX5X6X7X8KGGRHLIFCHSKKKCDEX9AX10X11LX12X13(K)n (SEQ ID NO:314), where X1is G, P, or S; X2is T, N, Q, H, or S; X3is E, T, or D; X4is K or R; X5is L or I; X6is E, A, S, or Q; X7is Q, T, Y, F, or L; X8is I or L; X9is L or I; X10is A, K, or S; X11is K, Q, or A; X12is T, R, or S; and X13is G or S, wherein n is an integer from 2 to 10, and wherein the TP42 T-cell epitope polypeptide has a length of from 34 amino acids to 52 amino acids; vii) a TP45 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2AVAX3YRGX4DVX5X6IPX7X8GDVVVX9X10TDALMTGX11TGDFDSVIDX12X13X14(K)n (SEQ ID NO:332), wherein X1is L or V; X2is N or T; X3is Y or F; X4is L or V; X5is S or A; X6is V or I; X7is T or A; X8is S, Q, or T; X9is V or C; X10is A or S; X11is F or Y; X12is C or K; X13is N or K; and X14is V or K, wherein n is an integer from 2 to 10, and wherein the TP45 T-cell epitope polypeptide has a length of from 36 amino acids to 55 amino acids; and viii) a TP48 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V, wherein n is an integer from 2 to 10, and wherein the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acids;ix) a TP33 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: HSKKKCDELAX1X2LX3X4X5GX6NAVAYYRGLDVSX7IP (SEQ ID NO:366), where X1is A or S; X2is K or A; X3is V, S, R, or T; X4is A or G; X5is L or M; X6is I, L, or V; and X7is V or I; wherein the TP33 T-cell epitope polypeptide has a length of from 30 amino acids to 36 amino acids; and x) a TP42-2 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KGGRHLIFCHSKKKCDELAX1X2LX3X4X5GX6NAVAYYRGLDVSX7IP (SEQ ID NO:371), where X1is A or S; X2is K or A; X3is V, S, R, or T; X4is A or G; X5is L or M; X6is I, L, or V; and X7is V or I; wherein the TP42-2 T-cell epitope polypeptide has a length of from 38 amino acids to 46 amino acids; xi) a TP240 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: SYQVGYLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRT VTTGAPITYSTYGKFLADGGCSGGAYDIIICDECHSX1DATTILGIGTVLDQAETAGARLVVLATA TPPGSVTVPHPNIEEVALGX2EGEIPFYGKAIPLX3X4IKGGRHLIFCHSKKKCDELAAKLRGMGLN AVAYYRGLDVSVIPTX5GDVVVVATDALMTGYTGDFDSVIDCNVAVTQT (SEQ ID NO:379), where X1is T, V, or Q, X2is T, N, Q, H, or S, X3is E, S, or A, X4is V, T, Y, F, or L, and X5is S, Q, or T; wherein the TP240 T-cell epitope polypeptide has a length of from 240 amino acids to 245 amino acids; xii) a TP65 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: TPIDTTIMAKNEVFCVDPX6KGGRKPARLIVYPDLGVRVCEKMALYDVVQKLPQAVMGX7SYGF QYS (SEQ ID NO:375), where X6is E, T, or V, and X7is S, A, or P; and wherein the TP65 T-cell epitope polypeptide has a length of from 65 amino acids to 70 amino acids; xiii) a TP156 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP156 amino acid sequence depicted in FIG.32C, where the TP156 T-cell epitope polypeptide has a length of from 150 amino acids to 160 amino acids; and xiv) a TP465 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 amino acid sequence depicted in any one of FIG.32B, 33B, and 34B, where the TP465 T-cell epitope polypeptide has a length of from 460 amino acids to 470 amino acids.

24. The composition of any one of claims 1-23, wherein the HCV E1 and / or E2 polypeptide is from HCV genotype 1, optionally genotype 1a.

25. The composition of any one of claims 1-23, wherein the HCV E1 and / or E2 polypeptide is from HCV genotype 2, optionally genotype 2a.

26. The composition of any one of claims 1-23, wherein the HCV E1 and / or E2 polypeptide is from HCV genotype 3, optionally genotype 3a.

27. The composition of any one of claims 1-23, wherein the HCV E1 and / or E2 polypeptide is / are from: a) HCV genotype 1, 2, and 3; b) HCV genotype 1a, 2, and 3; c) HCV genotype 1b, 2, and 3; or d) HCV genotype 1a, 1b, 2, and 3.

28. The composition of any one of claims 24-27, wherein the polypeptide-encoding region comprises a nucleotide sequence encoding an HCV Core polypeptide, wherein the HCV Core polypeptide is of genotype 1, genotype 2, or genotype 3.

29. The composition of any one of claims 17-28, wherein the one or more RNA molecules comprises a first RNA molecule, a second RNA molecule, and a third RNA molecule, wherein: a) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a first genotype; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; b) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a second genotype; ii) a second IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; and c) the polypeptide-encoding region of the third RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a third genotype; ii) a third IRES; andiii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; wherein the first IRES, the second IRES, and the third IRES are the same or different.

30. The composition of claim 29, wherein: a) the HCV E1 polypeptide encoded by the first RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the first RNA molecule is a soluble HCV E2 polypeptide; and / or b) the HCV E1 polypeptide encoded by the second RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the second RNA molecule is a soluble HCV E2 polypeptide; and / or c) the HCV E1 polypeptide encoded by the third RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the third RNA molecule is a soluble HCV E2 polypeptide.

31. The composition of claim 29 or claim 30, wherein the polypeptide-encoding region of one or more of the first RNA molecule, the second RNA molecule, and the third RNA molecule comprises a nucleotide sequence encoding an HCV Core polypeptide.

32. The composition of claim 29 or claim 30, wherein: a) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 1; ii) a first IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; b) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 2; ii) a second IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; and c) the polypeptide-encoding region of the third RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 3; ii) a third IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; wherein the first IRES, the second IRES, and the third IRES are the same or different.

33. The composition of claim 32, wherein:a) the HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E1 polypeptide, and the HCV E2 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E2 polypeptide; b) the HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E1 polypeptide, and the HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E2 polypeptide; and c) the HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E1 polypeptide, and the HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E2 polypeptide.

34. The composition of claim 32 or claim 33, wherein the polypeptide-encoding region of one or more of the first RNA molecule, the second RNA molecule, and the third RNA molecule comprises a nucleotide sequence encoding an HCV Core polypeptide.

35. The composition of claim 34, wherein: a) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polypeptide of genotype 1; ii) a first IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; b) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polypeptide of genotype 2; ii) a second IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; and c) the polypeptide-encoding region of the third RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polypeptide of genotype 3; ii) a third IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; wherein the first IRES, the second IRES, and the third IRES are the same or different.

36. The composition of claim 35, wherein: a) the HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E1 polypeptide, and the HCV E2polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E2 polypeptide; b) the HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 95% amino acid sequence identity to a J6 E1 polypeptide, and the HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E2 polypeptide; and c) the HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E1 polypeptide, and the HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E2 polypeptide.

37. The composition of any one of claims 17 to 28, wherein: a) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 1; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; b) the polypeptide-encoding region of the second RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 2; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; and c) the polypeptide-encoding region of the third RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 3; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

38. The composition of claim 37, wherein: a) the HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E1 polypeptide, and the HCV E2 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E2 polypeptide; b) the HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E1 polypeptide, and the HCV E2 polypeptideof genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E2 polypeptide; and c) the HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E1 polypeptide, and the HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E2 polypeptide.

39. The composition of claim 37, wherein: a) the HCV E1 polypeptide encoded by the first RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the first RNA molecule is a soluble HCV E2 polypeptide; and / or b) the HCV E1 polypeptide encoded by the second RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the second RNA molecule is a soluble HCV E2 polypeptide; and / or c) the HCV E1 polypeptide encoded by the third RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the third RNA molecule is a soluble HCV E2 polypeptide.

40. The composition of claim 39, wherein: a) the soluble HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble H77 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble H77 E2 polypeptide; b) the soluble HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble J6 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble J6 E2 polypeptide; and c) the soluble HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble S52 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble S52 E2 polypeptide.

41. The composition of any one of claims 37-40, wherein the polypeptide-encoding region of one or more of the first RNA molecule, the second RNA molecule, and the third RNA molecule comprises a nucleotide sequence encoding an HCV Core polypeptide.

42. The composition of claim 41, wherein: a) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 1; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; b) the polypeptide-encoding region of the second RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 2; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; and c) the polypeptide-encoding region of the third RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 3; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

43. The composition of claim 42, wherein: a) the HCV Core-E1-E2 polyprotein of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 Core-E1-E2 polyprotein; b) the HCV Core-E1-E2 polyprotein of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 Core-E1-E2 polyprotein; and c) the HCV Core-E1-E2 polyprotein of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 Core-E1-E2 polyprotein.

44. The composition of claim 41, wherein: a) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 1; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; b) the polypeptide-encoding region of the second RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 3; andii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; and c) the polypeptide-encoding region of the third RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 4, genotype 5, or genotype 6; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

45. The composition of any one of claims 23-44, wherein the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2 comprises: a) a TP50C T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGY P (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K, wherein the TP50C T-cell epitope polypeptide has a length of from 40 amino acids to 50 amino acids; b) a TP35-NS3 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), wherein X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R, wherein the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; c) a TP27 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S, wherein the TP27 T-cell epitope polypeptide has a length of from 22 amino acids to 27 amino acids; and d) a TP48 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T,or G; and X19is I, L, or V, wherein n is an integer from 2 to 10, and wherein the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acid.

46. The composition of any one of claims 23-45, wherein the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2 comprises, in order from N-terminus to C-terminus: a) a TP50C T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGY P (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K, wherein the TP50C T-cell epitope polypeptide has a length of from 40 amino acids to 50 amino acids; b) a TP35-NS3 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), wherein X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R, wherein the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; c) a TP27 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S, wherein the TP27 T-cell epitope polypeptide has a length of from 22 amino acids to 27 amino acids; and d) a TP48 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V, wherein n is an integer from 2 to 10, and wherein the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acid.

47. The composition of any one of claims 17-28, wherein the one or more RNA molecules comprises a first RNA molecule and a second RNA molecule, wherein:a) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding an HCV E1 and / or an HCV E2 polypeptide of a first genotype; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; and b) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding an HCV E1 and / or an HCV E2 polypeptide of a second genotype; ii) a second IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; wherein the first IRES and the second IRES are the same or different.

48. The composition of claim 47, wherein: a) the HCV E1 polypeptide encoded by the first RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the first RNA molecule is a soluble HCV E2 polypeptide; and / or b) the HCV E1 polypeptide encoded by the second RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the second RNA molecule is a soluble HCV E2 polypeptide.

49. The composition of claim 47 or 48, wherein the polypeptide-encoding region of one or both of the first RNA molecule and the second RNA molecule comprises a nucleotide sequence encoding an HCV Core polypeptide.

50. The composition of claim 47 or 48, wherein: a1) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 1; ii) a first IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; and b1) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 3; ii) a second IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; ora2) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 1; ii) a first IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; and b2) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 2; ii) a second IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; or a3) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 2; ii) a first IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; and b3) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 3; ii) a second IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; wherein the first IRES and the second IRES are the same or different; or a4) the first RNA molecule comprises. i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of a first genotype, optionally wherein the first genotype is genotype 1a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.32A; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP156 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP156 polypeptide amino acid sequence depicted in FIG.32C; and b4) the second RNA molecule comprises:i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a second genotype, optionally wherein the second genotype is genotype 2a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.34A; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP65 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP65 polypeptide amino acid sequence depicted in FIG.34C; or a5) the first RNA molecule comprises. i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of a first genotype, optionally wherein the first genotype is genotype 1a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.32A; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP156 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP156 polypeptide amino acid sequence depicted in FIG.32C; and b5) the second RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a second genotype, optionally wherein the second genotype is genotype 3a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.33A; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B;iv) a second IRES; and v) a nucleotide sequence encoding a TP48 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP48 polypeptide amino acid sequence depicted in FIG.33C; or a6) the first RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a first genotype, optionally wherein the first genotype is genotype 3a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.33A; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP48 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP48 polypeptide amino acid sequence depicted in FIG.33C; and b6) the second RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a second genotype, optionally wherein the second genotype is genotype 2a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.34A; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP65 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP65 polypeptide amino acid sequence depicted in FIG.34C.

51. The composition of claim 50, wherein: a) the HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E1 polypeptide, and the HCV E2polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E2 polypeptide; b) the HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E1 polypeptide, and the HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E2 polypeptide; and c) the HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E1 polypeptide, and the HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E2 polypeptide.

52. The composition of claim 50 or claim 51, wherein the polypeptide-encoding region of one or both of the first RNA molecule and the second RNA molecule comprises a nucleotide sequence encoding an HCV Core polypeptide.

53. The composition of claim 52, wherein: a1) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 1; ii) a first IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; b1) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 3; ii) a second IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; or a2) the polypeptide-encoding region of the first RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 1; ii) a first IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; b2) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 2; ii) a second IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; or a3) the polypeptide-encoding region of the first RNA molecule comprises:i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 2; ii) a first IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; b3) the polypeptide-encoding region of the second RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 3; ii) a second IRES; and iii) a nucleotide sequence encoding the fusion polypeptide; wherein the first IRES and the second IRES are the same or different.

54. The composition of claim 53, wherein: a) the HCV Core-E1-E2 polyprotein of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 HCV Core-E1-E2 polyprotein; b) the HCV Core-E1-E2 polyprotein of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 HCV Core-E1-E2 polyprotein; and c) the HCV Core-E1-E2 polyprotein of genotype 3 comprises an amino acid sequence at least 90%, or at least 95%, amino acid sequence identity to an S52 HCV Core-E1-E2 polyprotein.

55. The composition of any one of claims 17 to 28, wherein the composition comprises a first RNA molecule and a second RNA molecule, and wherein: a1) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 1; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; b1) the polypeptide-encoding region of the second RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 3; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or a2) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 1; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2;b2) the polypeptide-encoding region of the second RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 2; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or a3) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 2; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; b3) the polypeptide-encoding region of the second RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 3; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

56. The composition of claim 55, wherein: a) the HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E1 polypeptide, and the HCV E2 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E2 polypeptide; b) the HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E1 polypeptide, and the HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E2 polypeptide; and c) the HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E1 polypeptide, and the HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E2 polypeptide.

57. The composition of claim 55, wherein: a) the HCV E1 polypeptide encoded by the first RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the first RNA molecule is a soluble HCV E2 polypeptide; and / orb) the HCV E1 polypeptide encoded by the second RNA molecule is a soluble HCV E1 polypeptide and / or the HCV E2 polypeptide encoded by the second RNA molecule is a soluble HCV E2 polypeptide.

58. The composition of claim 57, wherein: a) the soluble HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble H77 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble H77 E2 polypeptide; b) the soluble HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble J6 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble J6 E2 polypeptide; and c) the soluble HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble S52 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble S52 E2 polypeptide.

59. The composition of any one of claims 55-58, wherein the polypeptide-encoding region of one or both of the first RNA molecule and the second RNA molecule comprises a nucleotide sequence encoding an HCV Core polypeptide.

60. The composition of claim 59, wherein: a1) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 1; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; b1) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 3; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; ora2) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 1; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; b2) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 2; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or a3) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 2; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; b3) the polypeptide-encoding region of the first RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 3; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

61. The composition of claim 60, wherein: a) the HCV Core-E1-E2 polyprotein of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 Core-E1-E2 polyprotein; b) the HCV Core-E1-E2 polyprotein of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 Core-E1-E2 polyprotein; and c) the HCV Core-E1-E2 polyprotein of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 Core-E1-E2 polyprotein.

62. The composition of any one of claims 47-61, wherein the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2 comprises: a) a TP50C T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to:GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGY P (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K, wherein the TP50C T-cell epitope polypeptide has a length of from 40 amino acids to 50 amino acids; b) a TP35-NS3 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), wherein X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R, wherein the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; c) a TP27 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S, wherein the TP27 T-cell epitope polypeptide has a length of from 22 amino acids to 27 amino acids; and d) a TP48 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V, wherein n is an integer from 2 to 10, and wherein the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acid.

63. The composition of any one of claims 47-61, wherein the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2 comprises, in order from N-terminus to C-terminus: a) a TP50C T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGY P (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K, wherein the TP50C T-cell epitope polypeptide has a length of from 40 amino acids to 50 amino acids;b) a TP35-NS3 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), wherein X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R, wherein the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; c) a TP27 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S, wherein the TP27 T-cell epitope polypeptide has a length of from 22 amino acids to 27 amino acids; and d) a TP48 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V, wherein n is an integer from 2 to 10, and wherein the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acid.

64. The composition of any one of claims 17-28, wherein the composition comprises a single type of RNA molecule, wherein: a) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV E1 and / or an HCV E2 polypeptide of genotype 1; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or b) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV E1 and / or an HCV E2 polypeptide of genotype 2; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; orc) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV E1 and / or an HCV E2 polypeptide of genotype 3; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or d) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV E1 and / or an HCV E2 polypeptide of genotype 4; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or e) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV E1 and / or an HCV E2 polypeptide of genotype 5; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or f) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV E1 and / or an HCV E2 polypeptide of genotype 6; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide.

65. The composition of claim 64, wherein: a) the HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E1 polypeptide, and the HCV E2 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E2 polypeptide;b) the HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, % amino acid sequence identity to a J6 E1 polypeptide, and the HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E2 polypeptide; and c) the HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E1 polypeptide, and the HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E2 polypeptide.

66. The composition of claim 64 or claim 65, wherein the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding an HCV Core polypeptide.

67. The composition of claim 66, wherein: a) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 1; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or b) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 2; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or c) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 3; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or d) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 4; ii) a first IRES; andiii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or e) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 5; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or f) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding an HCV Core-E1-E2 polyprotein of genotype 6; ii) a first IRES; and iii) one or more nucleotide sequences encoding the one or more heterologous polypeptides or the fusion polypeptide; or g) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 1a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.32A; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP156 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP156 polypeptide amino acid sequence depicted in FIG.32C; or h) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 3a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.33A; ii) a first IRES;iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP48 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP48 polypeptide amino acid sequence depicted in FIG.33C; or j) the polypeptide-encoding region of the RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of genotype 2a, optionally wherein the HCV E1 and E2 polypeptides comprise amino acid sequences having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.34A; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP65 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP65 polypeptide amino acid sequence depicted in FIG.34C.

68. The composition of claim 67, wherein: a) the HCV Core-E1-E2 polyprotein of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 HCV Core-E1-E2 polyprotein; b) the HCV Core-E1-E2 polyprotein of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 HCV Core-E1-E2 polyprotein; and c) the HCV Core-E1-E2 polyprotein of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 HCV Core-E1-E2 polyprotein.

69. The composition of any one of claims 17 to 28, wherein the composition comprises a single type of RNA molecule, and wherein: a) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 1; andii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or b) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 2; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or c) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 3; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or d) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 4; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or e) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 5; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or f) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) HCV E1 and HCV2 E2 polypeptides of genotype 6; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

70. The composition of claim 69, wherein:a) the HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E1 polypeptide, and the HCV E2 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 E2 polypeptide; b) the HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E1 polypeptide, and the HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 E2 polypeptide; and c) the HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E1 polypeptide, and the HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 E2 polypeptide.

71. The composition of claim 69, wherein: a) the HCV E1 polypeptide encoded by the RNA molecule is a soluble HCV E1 polypeptide; and / or b) the HCV E2 polypeptide encoded by the RNA molecule is a soluble HCV E2 polypeptide.

72. The composition of claim 71, wherein: a) the soluble HCV E1 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble H77 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble H77 E2 polypeptide; b) the soluble HCV E1 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble J6 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble J6 E2 polypeptide; and c) the soluble HCV E1 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble S52 E1 polypeptide, and the soluble HCV E2 polypeptide of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a soluble S52 E2 polypeptide.

73. The composition of any one of claims 69-72, wherein the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding an HCV Core polypeptide.

74. The composition of claim 73, wherein: a) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 1; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or b) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 2; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or c) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 3; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or d) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 4; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or e) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 5; and ii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2; or f) the polypeptide-encoding region of the RNA molecule comprises a nucleotide sequence encoding a fusion polypeptide comprising: i) an HCV Core-E1-E2 polyprotein of genotype 6; andii) the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

75. The composition of claim 74, wherein: a) the HCV Core-E1-E2 polyprotein of genotype 1 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an H77 Core-E1-E2 polyprotein; b) the HCV Core-E1-E2 polyprotein of genotype 2 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to a J6 Core-E1-E2 polyprotein; and c) the HCV Core-E1-E2 polyprotein of genotype 3 comprises an amino acid sequence having at least 90%, or at least 95%, amino acid sequence identity to an S52 Core-E1-E2 polyprotein.

76. The composition of any one of claims 64-75, wherein the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2 comprises: a) a TP50C T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGY P (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K, wherein the TP50C T-cell epitope polypeptide has a length of from 40 amino acids to 50 amino acids; b) a TP35-NS3 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), wherein X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R, wherein the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; c) a TP27 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S, wherein the TP27 T-cell epitope polypeptide has a length of from 22 amino acids to 27 amino acids; and d) a TP48 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V, wherein n is an integer from 2 to 10, and wherein the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acid.

77. The composition of any one of claims 64-75, wherein the fusion polypeptide comprising two or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2 comprises, in order from N-terminus to C-terminus: a) a TP50C T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: GVYX1LPRRGPRLGVRX2TRKX3SERSQPRGRRQX4IPKX5X6X7X8X9GX10X11WX12X13PGY P (SEQ ID NO:234), where X1is L or V; X2is A or G; X3is T or S; X4is P or R; X5is A or D; X6is R or A; X7is R, Q, or S; X8is S or P; X9is E, T, or Q; X10is R or K; X11is S, T, H, or A; X12is A or G; and X13is Q or K, wherein the TP50C T-cell epitope polypeptide has a length of from 40 amino acids to 50 amino acids; b) a TP35-NS3 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: KSTKVPX1AYX2X3QGYX4VLVLNPSVAATLGFGX5X6X7SX8(SEQ ID NO:219), wherein X1is A or V; X2is A or V; X3is A or S; X4is K or N; X5is A or S; X6is Y or F; X7is M or L; and X8is K or R, wherein the TP35-NS3 T-cell epitope polypeptide has a length of from 28 amino acids to 35 amino acids; c) a TP27 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3X4KGGRHLIFCHSKKKCDEX5AX6X7LX8(SEQ ID NO:273), where X1is L or I; X2is E, A, S, V, or Q; X3is Q, T, Y, F, or L; X4is I or L; X5is L or I; X6is A, K, or S; X7is K, Q, or A; and X8is T, R, or S, wherein the TP27 T-cell epitope polypeptide has a length of from 22 amino acids to 27 amino acids; and d) a TP48 T-cell epitope polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to: X1X2X3RHX4GX5X6EGX7X8QWMNRLIAFASRGNHVX9PTHYX10X11X12X13DAX14X15X16VX17X18X19L(K)n (SEQ ID NO:339), wherein X1is I or V; X2is L or I; X3is R or K; X4is V, I, or T; X5is P, Q, or T; X6is G, A, or S; X7is A or V; X8is V or T; X9is S or A; X10is V or I; X11is P, T, A, or Q, X12is E or D; X13is S, T, or D; X14is S or A; X15is A, Q, R, or K; X16is R, K, or X; X17is T or M; X18is Q, A, T, or G; and X19is I, L, or V, wherein n is an integer from 2 to 10, and wherein the TP48 T-cell epitope polypeptide has a length of from 38 amino acids to 58 amino acid.

78. The composition of any one of claims 17-28, wherein the one or more RNA molecules comprises a first RNA molecule, a second RNA molecule, and a third RNA molecule, wherein: a) the first RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of a first genotype; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP156 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP156 polypeptide amino acid sequence depicted in FIG.32C; b) the second RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a second genotype; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP48 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP48 polypeptide amino acid sequence depicted in FIG.33C; and c) the third RNA molecule comprises: i) a nucleotide sequence encoding HCV E1 and / or an HCV E2 polypeptide of a third genotype; ii) a first IRES; iii) a nucleotide sequence encoding a TP465 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP465 polypeptide amino acid sequence depicted in FIG.32B, FIG.33B, or FIG.34B; iv) a second IRES; and v) a nucleotide sequence encoding a TP65 polypeptide comprising an amino acid sequence having at least 80% amino acid sequence identity to the TP65 polypeptide amino acid sequence depicted in FIG.34C.

79. The composition of claim 78, wherein: a) the nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of the first genotype encodes HCV E1 and E2 polypeptides having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.32A; b) the nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of the second genotype encodes HCV E1 and E2 polypeptides having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.33A; and c) the nucleotide sequence encoding HCV E1 and HCV E2 polypeptides of the third genotype encodes HCV E1 and E2 polypeptides having at least 80% amino acid sequence identity to the E1 and E2 amino acid sequences depicted in FIG.34A.

80. The composition of any one of claims 1-79, wherein the RNA is a self-amplifying RNA comprising a nucleotide sequence encoding an RNA-dependent RNA polymerase (RdRP).

81. The composition of claim 80, wherein the self-amplifying RNA is a cis-replicon.

82. The composition of claim 80 or claim 81, wherein the self-amplifying RNA is a recombinant Venezuelan equine encephalitis virus, a Sindbis virus, or a recombinant Semliki Forest virus.

83. The composition of any one of claims 1-82, wherein the composition is in the form of a lipid nanoparticle.

84. The composition of any one of claims 1-82, wherein the composition is in the form of a liposome.

85. The composition of any one of claims 1-82, wherein the composition comprises one or more lipids selected from cationic lipids, neutral lipids, anionic lipids, helper lipids, and stealth lipids.

86. The composition of any one of claims 1-2, wherein the composition comprises an HCV Core polypeptide and wherein the one or more RNA molecules are within a virus-like particle comprising the HCV Core polypeptide.

87. A method of inducing an immune response to hepatitis C virus (HCV) in an individual, the method comprising administering to the individual an effective amount of a composition of any one of claims 1-86.

88. The method of claim 87, comprising: a) administering a first dose of the composition at a first time; and b) administering a second dose of the composition at a second time.

89. The method of claim 88, wherein the second time is from a period of time of from about 1 week to about 6 months from the first time.

90. The method of any one of claims 87-89, wherein said administering is intramuscular administration.

91. The method of any one of claims 87-89, wherein said administering is subcutaneous administration.

92. The method of any one of claims 87-91, comprising administering one or more heterologous polypeptides comprising a T-cell epitope present in an HCV protein other than E1 and E2.

93. The method of any one of claim 87-92, comprising administering an HCV E1 polypeptide and / or an HCV E2 polypeptide.

94. The method of claim 88, wherein: a) the first dose comprises one or more RNA molecules comprising a polypeptide-encoding region comprising one or more nucleotide sequences encoding a hepatitis C virus (HCV) E1 polypeptide and / or an HCV E2 polypeptide of a first genotype; and b) the second dose comprises one or more RNA molecules comprising a polypeptide-encoding region comprising one or more nucleotide sequences encoding a hepatitis C virus (HCV) E1 polypeptide and / or an HCV E2 polypeptide of a second genotype.

95. The method of claim 94, wherein: a) the first dose comprises a composition of any one of claims 64-76; and b) the second dose comprises a composition of any one of claims 64-76.

96. The method of claim 94, wherein: a1) the first dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 1; and b1) the second dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 3; or a2) the first dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 1; and b2) the second dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 4; or a3) the first dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 1; and b3) the second dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 5; or a4) the first dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 1; and b4) the second dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 6; or a5) the first dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 1; and b5) the second dose comprises a composition of any one of claims 64-76, wherein the HCV E1 and / or HCV E2 polypeptide are of genotype 2.

97. The method of any one of claim 88-94, comprising administering at least a third dose of the composition at a third time.

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