RNA compositions encoding herpes simplex virus glycoprotein b antigens and uses thereof

Abstract

The present disclosure provides a polyribonucleotide encoding a polypeptide that comprises a viral antigen from Herpes Simplex Virus-2 (HSV-2); for example, the viral antigen comprises the ectodomain of HSV-2 glycoprotein B. The polyribonucleotide can be formulated as an RNA composition useful for inducing anti-HSV immune responses in a subject.

Description

P-628703-USP RNA COMPOSITIONS ENCODING HERPES SIMPLEX VIRUS GLYCOPROTEIN B ANTIGENS AND USES THEREOF SEQUENCE LISTING STATEMENT

[0001] The instant application contains a Sequence Listing conforming to the rules of WIPO Standard ST.26 which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. The XML copy, created on August 2, 2024, is titled P-682703-PC-SEQLIST2.xml and is 415 kilobytes in size. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with U.S. government support under AI139618 awarded by The National Institutes of Health. The government has certain rights in the invention. TECHNICAL FIELD

[0003] The present disclosure relates in general to the field of RNA therapeutics. In some embodiments, the present disclosure provides RNA constructs encoding HSV-2 glycoprotein as immunotherapy for genital herpes. BACKGROUND

[0004] A half-billion people worldwide are infected with herpes simplex virus type 2 (HSV-2). Many of these individuals are unaware they are infected, yet they are at risk of transmitting infection to intimate partners. About 20% of infected people have frequent, painful recurrent genital lesions. Lifelong daily suppressive therapy with acyclovir or valacyclovir reduces the frequency of recurrences and lowers risk for transmission, but not all people respond or are willing to take daily therapy. Anxiety about transmission to intimate partners is perhaps the greatest concern of people with genital herpes.

[0005] One of the most dreaded complications of genital herpes is neonatal herpes. This infection is uncommon (1:3,000 births in the U.S.) but devastating with high morbidity and mortality in newborns. Neonates acquire herpes simplex virus type 1 (HSV-1) or HSV-2 infection from mothers who have reactivation infection at the time of labor and delivery, or the infection in the pregnant woman can be a first-time infection late in pregnancy.

[0006] Prophylactic vaccines that are under development are intended to prevent first-time HSV infections, and those vaccines may not be effective in preventing recurrences in people already infected. From a public health perspective, the biggest impact of an effective genital herpes vaccine will be on HIV infection. Genital herpes increases the risk of acquiring or transmitting HIV by 3-4-fold. SUMMARY

[0007] In some embodiments, described herein is a polyribonucleotide (e.g., a nucleoside-modified polyribonucleotide) encoding a polypeptide, wherein the polypeptide comprises the ectodomain of a Herpes Simplex Virus-2 (HSV-2) glycoprotein B (gB) antigen or an immunogenic fragment thereof. In some embodiments, the ectodomain of the HSV-2 gB comprises the amino acid sequence as set forth in SEQ ID NO: 1. In some Page 1 of 182 12197519v1embodiments, a polyribonucleotide disclosed herein comprises the ribonucleic acid sequence as set forth in SEQ ID NO: 5.

[0008] In some embodiments, a polyribonucleotide disclosed herein further comprises a secretory signal sequence encoding an IL2 secretory signal polypeptide; for example, an IL2 secretory signal polypeptide having the amino acid sequence as set forth in SEQ ID NO: 2.

[0009] In some embodiments, a polyribonucleotide disclosed herein further comprises a secretory signal sequence encoding an HSV gB secretory signal polypeptide; for example, an HSV gB secretory signal polypeptide having the amino acid sequence as set forth in SEQ ID NO: 4.

[0010] In some embodiments, a polyribonucleotide disclosed herein further comprises a 5′ untranslated region. In some embodiments, a polyribonucleotide disclosed herein further comprises a 3′ untranslated region. In some embodiments, a polyribonucleotide disclosed herein further comprises a poly-A tail.

[0011] In some embodiments, a polyribonucleotide disclosed herein comprises the ribonucleic acid sequence as set forth in SEQ ID NO: 6.

[0012] In another embodiment, the present disclosure provides a composition or combination comprising a polyribonucleotide disclosed herein. In some embodiments, the composition or combination comprises a nanoparticle, lipid, polymer, cholesterol, or cell penetrating peptide that encapsulates the polyribonucleotide.

[0013] In some embodiments, the composition or combination further comprises one or more RNAs encoding: a) HSV glycoprotein C (gC) or an immunogenic fragment thereof, b) HSV glycoprotein D (gD) or an immunogenic fragment thereof, c) HSV glycoprotein E (gE) or an immunogenic fragment thereof, d) HSV glycoprotein H (gH) or an immunogenic fragment thereof, e) HSV glycoprotein L (gL) or an immunogenic fragment thereof, f) HSV glycoprotein I (gI) or an immunogenic fragment thereof, or g) any combination thereof.

[0014] In another embodiment, there is provided a method of using a polyribonucleotide disclosed herein, or a composition or combination disclosed herein, to treat a Herpes Simplex Virus (HSV) infection in a subject.

[0015] In another embodiment, there is provided a method of using a polyribonucleotide disclosed herein, or the composition or combination disclosed herein, to suppress, inhibit, or reduce the incidence of a Herpes Simplex Virus (HSV) infection in a subject.

[0016] In another embodiment, there is provided a method of using the polyribonucleotide disclosed herein, or the composition or combination disclosed herein, to induce an anti-HSV immune response in a subject.

[0017] These and other aspects of the invention will be appreciated from the ensuing descriptions of the figures and detailed description of the invention. BRIEF DESCRIPTION OF THE DRAWINGSPage 2 of 182 12197519v1

[0018] Some embodiments of the invention are herein described, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of embodiments of the invention. In this regard, the description taken with the drawings makes apparent to those skilled in the art how embodiments of the invention may be practiced.

[0019] Figure 1 shows cumulative recurrent genital lesion days per group. Previously infected guinea pigs were immunized on days 35 and 65 post-infection with nucleoside-modified RNA encapsulated in lipid nanoparticle 315 and expressing an exemplary immunogenic fragment of HSV-2 gB (gB2) (30 μg) or PBS (control). Animals were scored daily Monday to Friday for recurrent genital lesions from 1 day after the first immunization until the end of the study on day 116. From the time of the second immunization, major differences appeared comparing the gB2 group with the PBS group.

[0020] Figure 2 shows days with recurrent genital lesions for each animal starting 1 day after the second immunization. The same animals as in Figure 1 are shown here. P values were calculated by the two-tailed Mann Whitney test and demonstrate highly significant differences comparing the gB2 group with PBS (**, P<0.01).

[0021] Figure 3 shows cytokine production by CD4+ T cell in response to compositions comprising HSV-2 glycoproteins. Mice were immunized twice as described herein with 10 µg of an exemplary immunogenic fragment of HSV-2 gB (gB2) RNA-LNP. Splenocytes from these mice were stimulated with a gB2 overlapping peptide pool. CD4+ cytokine-producing T cells were analyzed by flow cytometry.

[0022] Figure 4 shows cytokine production by CD8+ T cell in response to compositions comprising HSV-2 glycoproteins. Mice were immunized twice as described herein with 10 µg of an exemplary immunogenic fragment of HSV-2 gB (gB2) RNA-LNP. Splenocytes from these mice were stimulated with a gB2 overlapping peptide pool. CD8+ cytokine-producing T cells were analyzed by flow cytometry.

[0023] Figure 5 shows expression levels in HEK293T cells transfected with RNA encoding HSV-2 gB (gB2) antigens. Constructs encoding gB2 antigens include 4059 (SEQ ID NO: 219), 4060 (SEQ ID NO: 220), 4061 (SEQ ID NO: 221), 4063 (SEQ ID NO: 225), 4064 (SEQ ID NO: 226), and 4065 (SEQ ID NO: 227). Representative data from one experiment showing median fluorescence intensities (MFI) of the total HEK293T population for gB2 antigen constructs. Data shown are mean+SD of HEK293T transfections performed in triplicates.

[0024] Figures 6A-6D show the prophylactic effect of BNT163, gB, gE2 / gI2, or a combination of them on survival (Figure 6A), disease severity (Figure 6B), genital lesions (Figure 6C), and urinary retention (Figure 6D) in guinea pigs infected with HSV-2. As used in Figures 6A-6D, “BNT163” refers to a combination of three polyribonucleotides: (i) a polyribonucleotide encoding a polypeptide comprising a gC immunogenic fragment having an amino acid sequence according SEQ ID NO: 258; (ii) a polyribonucleotide encoding a polypeptide comprising a gD immunogenic fragment having an amino acid sequence according SEQ ID NO: 260; and (iii) a polyribonucleotide encoding a polypeptide comprising a gE immunogenic fragment having an amino acid sequence according SEQ ID NO: 262.

[0025] Figures 7A-7B show the therapeutic effect of BNT163, gB and gE2 / gI2 on recurrent lesions in a first (Figure 7A) and the combined results of two experiments (Figure 7B) experiments. As used in Figures 7A-7B,Page 3 of 182 12197519v1“BNT163” refers to a combination of three polyribonucleotides: (i) a polyribonucleotide encoding a polypeptide comprising a gC immunogenic fragment having an amino acid sequence according SEQ ID NO: 258; (ii) a polyribonucleotide encoding a polypeptide comprising a gD immunogenic fragment having an amino acid sequence according SEQ ID NO: 260; and (iii) a polyribonucleotide encoding a polypeptide comprising a gE immunogenic fragment having an amino acid sequence according SEQ ID NO: 262.

[0026] Figures 8A-8B show T cell responses to immunization with gB2 RNA in CD4+ (Figure 8A) and CD8+ (Figure 8B) T cells. DETAILED DESCRIPTION

[0027] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and / or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting. Each literature reference or other citation referred to herein is incorporated herein by reference in its entirety.

[0028] In the description presented herein, each of the steps of the invention and variations thereof are described. This description is not intended to be limiting and changes in the components, sequence of steps, and other variations would be understood to be within the scope of the present invention.

[0029] 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 subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements. Definitions

[0030] As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise.

[0031] As used herein, the term “about”, when used herein in reference to a value, refers to a value that is similar, in context to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by “about” in that context. For example, in some embodiments, the term “about” may encompass a range of values that within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.

[0032] As used herein, the term “adjuvant” refers to compounds that, when administered to an individual or tested in vitro, increase the immune response to an antigen in the individual or test system to which the antigen is administered. In some embodiments, an immune adjuvant enhances an immune response to an antigen that isPage 4 of 182 12197519v1weakly immunogenic when administered alone, i.e., inducing no or weak antibody titers or cell-mediated immune response. In some embodiments, the adjuvant increases antibody titers to the antigen. In some embodiments, the adjuvant lowers the dose of the antigen effective to achieve an immune response in the individual. Multiple types of adjuvants are known in the art and described in detail in U.S. Patent Publication 2013 / 0028925 which is hereby incorporated by reference herein.

[0033] As used herein, the term “administering” refers to directly introducing into a subject by injection or other means a composition of the present disclosure. In another embodiment, “administering” refers to contacting a cell of the subject’s immune system with a composition or modified polyribonucleotides encoding HSV gE and gI.

[0034] As used herein, the term “agent” may refer to a physical entity or phenomenon. In some embodiments, an agent may be characterized by a particular feature and / or effect. In some embodiments, an agent may be a compound, molecule, or entity of any chemical class including, for example, a small molecule, polypeptide, nucleic acid, saccharide, lipid, metal, or a combination or complex thereof. In some embodiments, the term “agent” may refer to a compound, molecule, or entity that comprises a polymer. In some embodiments, the term may refer to a compound or entity that comprises one or more polymeric moieties. In some embodiments, the term “agent” may refer to a compound, molecule, or entity that is substantially free of a particular polymer or polymeric moiety. In some embodiments, the term may refer to a compound, molecule, or entity that lacks or is substantially free of any polymer or polymeric moiety.

[0035] As used herein, the term “amino acid,” in its broadest sense, refers to a compound and / or substance that can be, is, or has been incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N–C(H)(R)–COOH. In some embodiments, an amino acid is a naturally-occurring amino acid. In some embodiments, an amino acid is a non-natural amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L-amino acid. “Standard amino acid” refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. “Nonstandard amino acid” refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. In some embodiments, an amino acid, including a carboxy- and / or amino-terminal amino acid in a polypeptide, can contain a structural modification as compared with the general structure above. For example, in some embodiments, an amino acid may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and / or the hydroxyl group) as compared with the general structure. In some embodiments, such modification may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino acid. In some embodiments, such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino acid, as compared with one containing an otherwise identical unmodified amino acid. As will be clear from context, in some embodiments, the term “amino acid” may be used to refer to a free amino acid; in some embodiments it may be used to refer to an amino acid residue of a polypeptide.

[0036] As used herein, the term “antibody agent” refers to an agent that specifically binds to a particular antigen. In some embodiments, the term encompasses a polypeptide or polypeptide complex that includes immunoglobulin structural elements sufficient to confer specific binding. For example, in some embodiments, an antibody agent is orPage 5 of 182 12197519v1comprises a polypeptide whose amino acid sequence includes one or more structural elements recognized by those skilled in the art as a complementarity determining region (CDR); in some embodiments an antibody agent is or comprises a polypeptide whose amino acid sequence includes at least one CDR (e.g., at least one heavy chain CDR and / or at least one light chain CDR) that is substantially identical to one found in a reference antibody. In some embodiments an included CDR is substantially identical to a reference CDR in that it is either identical in sequence or contains between 1-5 amino acid substitutions as compared with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that it shows at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that it shows at least 96%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR is deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR is substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments, an antibody agent is or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art as an immunoglobulin variable domain. In some embodiments, an antibody agent in or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art to correspond to CDRs1, 2, and 3 of an antibody variable domain; in some such embodiments, an antibody agent in or comprises a polypeptide or set of polypeptides whose amino acid sequence(s) together include structural elements recognized by those skilled in the art to correspond to both heavy chain and light chain variable region CDRs, e.g., heavy chain CDRs 1, 2, and / or 3 and light chain CDRs 1, 2, and / or 3. In some embodiments, an antibody agent is a polypeptide protein having a binding domain which is homologous or largely homologous to an immunoglobulin-binding domain. In some embodiments, an antibody agent may be or comprise a polyclonal antibody preparation. In some embodiments, an antibody agent may be or comprise a monoclonal antibody preparation. In some embodiments, an antibody agent may include one or more constant region sequences that are characteristic of a particular organism, such as a camel, human, mouse, primate, rabbit, rat; in many embodiments, an antibody agent may include one or more constant region sequences that are characteristic of a human. In some embodiments, an antibody agent may include one or more sequence elements that would be recognized by one skilled in the art as a humanized sequence, a primatized sequence, a chimeric sequence, etc. In some embodiments, an antibody agent may be a canonical antibody (e.g., may comprise two heavy chains and two light chains). In some embodiments, an antibody agent may be in a format selected from, but not limited to, intact IgA, IgG, IgE or IgM antibodies; bi- or multi- specific antibodies (e.g., Zybodies®, etc); antibody fragments such as Fab fragments, Fab’ fragments, F(ab’)2 fragments, Fd’ fragments, Fd fragments, and isolated CDRs or sets thereof; single chain Fvs; polypeptide-Fc fusions; single domain antibodies (e.g., shark singlePage 6 of 182 12197519v1domain antibodies such as IgNAR or fragments thereof); cameloid antibodies; masked antibodies (e.g., Probodies®); Small Modular ImmunoPharmaceuticals (“SMIPsTM”); single chain or Tandem diabodies (TandAb®); VHHs; Anticalins®; Nanobodies® minibodies; BiTE®s; ankyrin repeat proteins or DARPINs®; Avimers®; DARTs; TCR-like antibodies;, Adnectins®; Affilins®; Trans-bodies®; Affibodies®; TrimerX®; MicroProteins; Fynomers®, Centyrins®; and KALBITOR®s. In some embodiments, an antibody may lack a covalent modification (e.g., attachment of a glycan) that it would have if produced naturally. In some embodiments, an antibody may contain a covalent modification (e.g., attachment of a glycan, a payload (e.g., a detectable moiety, a therapeutic moiety, a catalytic moiety, etc.), or other pendant group (e.g., poly-ethylene glycol, etc.)).

[0037] As used herein, the term “antigen” refers to a molecule that is recognized by the immune system, e.g., in particular embodiments the adaptive immune system, such that it elicits an antigen-specific immune response. In some embodiments, an antigen-specific immune response may be or comprise generation of antibodies and / or antigen-specific T cells. In some embodiments, an antigen is a peptide or polypeptide that comprises at least one epitope against which an immune response can be generated. In some embodiments, an antigen is presented by cells of the immune system such as antigen presenting cells like dendritic cells or macrophages. In one embodiments, an antigen or a processed product thereof such as a T-cell epitope is bound by a T- or B-cell receptor, or by an immunoglobulin molecule such as an antibody. Accordingly, an antigen or a processed product thereof may react specifically with antibodies or T lymphocytes (T cells). In some embodiments, an antigen is a parasitic antigen. In accordance with the present disclosure, in some embodiments, an antigen may be delivered by RNA molecules as described herein. In some embodiments, a peptide or polypeptide antigen can be 2-100 amino acids, including for example, 5 amino acids, 10 amino acids, 15 amino acids, 20 amino acids, 25 amino acids, 30 amino acids, 35 amino acids, 40 amino acids, 45 amino acids, or 50 amino acids in length. In some embodiments, a peptide or polypeptide antigen can be greater than 50 amino acids. In some embodiments, a peptide or polypeptide antigen can be greater than 100 amino acids. In some embodiments, an antigen is recognized by an immune effector cell. In some embodiments, an antigen if recognized by an immune effector cell is able to induce in the presence of appropriate co-stimulatory signals, stimulation, priming and / or expansion of the immune effector cell carrying an antigen receptor recognizing the antigen. In the context of the embodiments of the present disclosure, in some embodiments, an antigen can be presented or present on the surface of a cell, e.g., an antigen presenting cell. In some embodiments, an antigen is presented by a diseased cell such as a virus-infected cell. In some embodiments, an antigen receptor is a TCR which binds to an epitope of an antigen presented in the context of MHC. In some embodiments, binding of a TCR when expressed by T cells and / or present on T cells to an antigen presented by cells such as antigen presenting cells results in stimulation, priming and / or expansion of said T cells. In some embodiments, binding of a TCR when expressed by T cells and / or present on T cells to an antigen presented on diseased cells results in cytolysis and / or apoptosis of the diseased cells, wherein said T cells preferably release cytotoxic factors, e.g., perforins and granzymes.

[0038] As used herein, the term "antigenic" refers to a peptide capable of specifically interacting with an antigen recognition molecule of the immune system, e.g. an immunoglobulin (antibody) or T cell antigen receptor.

[0039] As used herein, two events or entities are “associated” with one another, as that term is used herein, if the presence, level, degree, type and / or form of one is correlated with that of the other. For example, a particularPage 7 of 182 12197519v1entity (e.g., polypeptide, genetic signature, metabolite, microbe, etc) is considered to be associated with a particular disease, disorder, or condition, if its presence, level and / or form correlates with incidence of, susceptibility to, severity of, stage of, etc. the disease, disorder, or condition (e.g., across a relevant population). In some embodiments, two or more entities are physically “associated” with one another if they interact, directly or indirectly, so that they are and / or remain in physical proximity with one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non- covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof.

[0040] As used herein, that the term “binding” typically refers to a non-covalent association between or among entities or moieties. In some embodiments, binding data are expressed in terms of “IC50”. As is understood in the art, IC50 is the concentration of an assessed agent in a binding assay at which 50% inhibition of binding of reference agent known to bind the relevant binding partner is observed. In some embodiments, assays are run under conditions in which the assays are run (e.g., limiting binding target and reference concentrations), these values approximate KD values. Assays for determining binding are well known in the art and are described in detail, for example, in PCT publications WO 94 / 20127 and WO 94 / 03205, and other publications such Sidney et al., Current Protocols in Immunology 18.3.1 (1998); Sidney, et al., J. Immunol. 154:247 (1995); and Sette, et al., Mol. Immunol. 31:813 (1994). Alternatively, binding can be expressed relative to binding by a reference standard peptide. For example, can be based on its IC50, relative to the IC50of a reference standard peptide. Binding can also be determined using other assay systems including those using: live cells (e.g., Ceppellini et al., Nature 339:392 (1989); Christnick et al., Nature 352:67 (1991); Busch et al., Int. Immunol. 2:443 (1990); Hill et al., J. Immunol. 147:189 (1991); del Guercio et al., J. Immunol. 154:685 (1995)), cell free systems using detergent lysates (e.g., Cerundolo et al., J. Immunol 21:2069 (1991)), immobilized purified MHC (e.g., Hill et al., J. Immunol. 152, 2890 (1994); Marshall et al., J. Immunol. 152:4946 (1994)), ELISA systems (e.g., Reay et al., EMBO J. 11:2829 (1992)), surface plasmon resonance (e.g., Khilko et al., J. Biol. Chem. 268:15425 (1993)); high flux soluble phase assays (Hammer et al., J. Exp. Med. 180:2353 (1994)), and measurement of class I MHC stabilization or assembly (e.g., Ljunggren et al., Nature 346:476 (1990); Schumacher et al., Cell 62:563 (1990); Townsend et al., Cell 62:285 (1990); Parker et al., J. Immunol. 149:1896 (1992)).

[0041] As used herein, the term “C5-interfering domain” refers to a domain that interferes with binding of a host C3b molecule with a host C5 molecule. In another embodiment, the term refers to a domain that interferes with the interaction of a host C3b molecule with a host C5 molecule.

[0042] As used herein, the term “cap” refers to a structure comprising or essentially consisting of a nucleoside-5 '-triphosphate that is typically joined to a 5'-end of an uncapped RNA (e.g., an uncapped RNA having a 5'- diphosphate). In some embodiments, a cap is or comprises a guanine nucleotide. In some embodiments, a cap is or comprises a naturally-occurring RNA 5’ cap, including, e.g., but not limited to a 7- methylguanosine cap, which has a structure designated as “m7G.” In some embodiments, a cap is or comprises a synthetic cap analog that resembles an RNA cap structure and possesses the ability to stabilize RNA if attached thereto, including, e.g., but not limited to anti-reverse cap analogs (ARCAs) known in the art). Those skilled in the art will appreciate that methods for joining aPage 8 of 182 12197519v1cap to a 5’ end of an RNA are known in the art. For example, in some embodiments, a capped RNA may be obtained by in vitro capping of RNA that has a 5' triphosphate group or RNA that has a 5' diphosphate group with a capping enzyme system (including, e.g., but not limited to vaccinia capping enzyme system or Saccharomyces cerevisiae capping enzyme system). Alternatively, a capped RNA can be obtained by in vitro transcription (IVT) of a single- stranded DNA template in the presence of a dinucleotide or trinucleotide cap analog.

[0043] As used herein, the term “co-administration” refers to use of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent. The combined use of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent may be performed concurrently or separately (e.g., sequentially in any order). In some embodiments, a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent may be combined in one pharmaceutically-acceptable carrier, or they may be placed in separate carriers and delivered to a target cell or administered to a subject at different times. Each of these situations is contemplated as falling within the meaning of “co-administration” or “combination,” provided that a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent are delivered or administered sufficiently close in time that there is at least some temporal overlap in biological effect(s) generated by each on a target cell or a subject being treated.

[0044] As used herein, the term “codon-optimized” refers to alteration of codons in a coding region of a nucleic acid molecule to reflect the typical codon usage of a host organism. In some embodiments, codon optimization does not alter the amino acid sequence encoded by the nucleic acid molecule. Within the context of the present disclosure, in some embodiments coding regions are codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein. In some embodiments, codon-optimization may be performed such that codons for which frequently occurring tRNAs are available are inserted in place of “rare codons.” In some embodiments, codon- optimization may include increasing guanosine / cytosine (G / C) content of a coding region of RNA described herein as compared to the G / C content of the corresponding coding sequence of a wild type RNA, wherein the amino acid sequence encoded by the RNA is not modified compared to the amino acid sequence.

[0045] As used herein, the term “combination” refers to two or more agents (e.g., two or more polyribonucleotides) or compositions (e.g., therapeutic compositions) that are used together. In some embodiments, two or more agents (e.g., two or more polyribonucleotides) or compositions (e.g., therapeutic compositions) are used (e.g., manufactured, formulated, sold, or administered) together. For example, in some embodiments, two or more agents or compositions may be used (e.g., manufactured, formulated, sold, or administered) as part of the same treatment or prevention regimen. In some embodiments, two or more agents or compositions may be administered simultaneously or near-simultaneously (e.g., within 24 hours or less of each other, within 12 hours or less of each other, within 6 hours or less of each other, within 3 hours or less of each other, within 2 hours or less of each other, within 1 hour or less of each other, within 30 minutes or less of each other, within 15 minutes or less of each other, within 5 minutes or less of each other, or within 1 minute or less of each other) to a subject. For clarity, a combination does not require that individual agents or compositions be used (e.g., manufactured, formulated, sold, or administered) together in a single composition; although in some embodiments, two or more agents or compositions may be used (e.g., manufactured, formulated, sold, or administered) together in a single composition.Page 9 of 182 12197519v1

[0046] As used herein, the term “combination therapy” refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents). In some embodiments, the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all “doses” of a first regimen are administered prior to administration of any doses of a second regimen); in some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, “administration” of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the combination. For clarity, combination therapy does not require that individual agents be administered together in a single composition (or even necessarily at the same time), although in some embodiments, two or more agents, or active moieties thereof, may be administered together in a combination composition.

[0047] As used herein, the term “comparable” refers to two or more agents, entities, situations, sets of conditions, etc., that may not be identical to one another but that are sufficiently similar to permit comparison there between so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed. In some embodiments, comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features. Those of ordinary skill in the art will understand, in context, what degree of identity is required in any given circumstance for two or more such agents, entities, situations, sets of conditions, etc. to be considered comparable. For example, those of ordinary skill in the art will appreciate that sets of circumstances, individuals, or populations are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicative of the variation in those features that are varied.

[0048] As used herein, the terms “comprise”, "comprises", "comprising", "includes", "including", “having” and their conjugates mean "including but not limited to".

[0049] As used herein, the term “consisting of” means “including and limited to”. The term "consisting essentially of" means that the composition, method or structure may include additional ingredients, steps and / or parts, but only if the additional ingredients, steps and / or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.

[0050] As used herein, the term "consisting essentially of" means that the composition, method or structure may include additional ingredients, steps and / or parts, but only if the additional ingredients, steps and / or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.

[0051] As used herein, the term “corresponding to” refers to a relationship between two or more entities. For example, the term “corresponding to” may be used to designate the position / identity of a structural element in a compound or composition relative to another compound or composition (e.g., to an appropriate reference compound or composition). For example, in some embodiments, a monomeric residue in a polymer (e.g., an amino acid residue in a polypeptide or a nucleic acid residue in a polynucleotide) may be identified as “corresponding to” a residue in an appropriate reference polymer. For example, those of ordinary skill will appreciate that, for purposes of simplicity, residues in a polypeptide are often designated using a canonical numbering system based on a reference relatedPage 10 of 182 12197519v1polypeptide, so that an amino acid “corresponding to” a residue at position 190, for example, need not actually be the 190thamino acid in a particular amino acid chain but rather corresponds to the residue found at 190 in the reference polypeptide; those of ordinary skill in the art readily appreciate how to identify “corresponding” amino acids. For example, those skilled in the art will be aware of various sequence alignment strategies, including software programs such as, for example, BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH / GLSEARCH, Genoogle, HMMER, HHpred / HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE that can be utilized, for example, to identify “corresponding” residues in polypeptides and / or nucleic acids in accordance with the present disclosure. Those of skill in the art will also appreciate that, in some instances, the term “corresponding to” may be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity). To give but one example, a gene or protein in one organism may be described as “corresponding to” a gene or protein from another organism in order to indicate, in some embodiments, that it plays an analogous role or performs an analogous function and / or that it shows a particular degree of sequence identity or homology, or shares a particular characteristic sequence element.

[0052] As used herein, in the context of an amino acid sequence (peptide or polypeptide) “derived from” a designated amino acid sequence (peptide or polypeptide), refers to a structural analogue of a designated amino acid sequence. In some embodiments, an amino acid sequence which is derived from a particular amino acid sequence has an amino acid sequence that is identical, essentially identical or homologous to that particular sequence or a fragment thereof. Amino acid sequences derived from a particular amino acid sequence may be variants of that particular sequence or a fragment thereof. For example, it will be understood by one of ordinary skill in the art that the antigens suitable for use herein may be altered such that they vary in sequence from the naturally occurring or native sequences from which they were derived, while retaining the desirable activity of the native sequences.

[0053] As used herein, the term “designed” refers to an agent (i) whose structure is or was selected by the hand of man; (ii) that is produced by a process requiring the hand of man; and / or (iii) that is distinct from natural substances and other known agents.

[0054] As used herein, the term “dosing regimen” may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimen comprises a plurality of doses each of which is separated in time from other doses. In some embodiments, individual doses are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).Page 11 of 182 12197519v1

[0055] As used herein, “encoding” refers to an RNA molecule that contains a gene that encodes a protein of interest, or a fragment thereof. In another embodiment, an RNA molecule comprises or consists of a protein coding sequence that encodes a protein of interest, or a fragment thereof. In another embodiment, one or more other proteins, or a fragment thereof is also encoded. In another embodiment, the protein of interest, or a fragment thereof, is the only protein encoded. Each possibility represents a separate embodiment of the present disclosure.

[0056] As used herein, the term “engineered” refers to the aspect of having been manipulated by the hand of man. For example, a polynucleotide is considered to be “engineered” when two or more sequences that are not linked together in that order in nature are manipulated by the hand of man to be directly linked to one another in the engineered polynucleotide and / or when a particular residue in a polynucleotide is non-naturally occurring and / or is caused through action of the hand of man to be linked with an entity or moiety with which it is not linked in nature.

[0057] As used herein, the term “epitope” refers to a moiety that is specifically recognized by an immunoglobulin (e.g., antibody or receptor) binding component. For example, an epitope may be recognized by a T cell, a B cell, or an antibody. In some embodiments, an epitope is comprised of a plurality of chemical atoms or groups on an antigen. In some embodiments, such chemical atoms or groups are surface-exposed when the antigen adopts a relevant three-dimensional conformation. In some embodiments, such chemical atoms or groups are physically near to each other in space when the antigen adopts such a conformation. In some embodiments, at least some such chemical atoms are groups are physically separated from one another when the antigen adopts an alternative conformation (e.g., is linearized). Accordingly, in some embodiments, an epitope of an antigen may include a continuous or discontinuous portion of the antigen. In some embodiments, an epitope is or comprises a T cell epitope. In some embodiments, an epitope may have a length of about 5 to about 30 amino acids, or about 10 to about 25 amino acids, or about 5 to about 15 amino acids, or about 5 to 12 amino acids, or about 6 to about 9 amino acids.

[0058] As used herein, the term “expression” of a nucleic acid sequence refers to the generation of a gene product from the nucleic acid sequence. In some embodiments, a gene product can be a transcript. In some embodiments, a gene product can be a polypeptide. In some embodiments, expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, etc); (3) translation of an RNA into a polypeptide or protein; and / or (4) post-translational modification of a polypeptide or protein.

[0059] As used herein, the terms “five prime untranslated region” or “5' UTR” refer to a sequence of an RNA molecule between a transcription start site and a start codon of a coding region of an RNA. In some embodiments, “5’ UTR” refers to a sequence of an RNA molecule that begins at a transcription start site and ends one nucleotide (nt) before a start codon (usually AUG) of a coding region of an RNA molecule, e.g., in its natural context.

[0060] As used herein, the term "fragment" refers to a protein or polypeptide that is shorter or comprises fewer amino acids than the full-length protein or polypeptide. In another embodiment, fragment refers to a nucleic acid encoding the protein fragment that is shorter or comprises fewer nucleotides than the full-length nucleic acid. In one embodiment, the fragment is an N-terminal fragment. In another embodiment, the fragment is a C-terminal fragment. In one embodiment, the fragment of the HSV protein is an ectodomain of the protein. In another embodiment, the fragment is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids shorter than the full-length protein. InPage 12 of 182 12197519v1another embodiment, the fragment is 50-100, 100-150, 150-300, or 300-600 amino acids shorter than the full-length protein. In another embodiment, the fragment comprises approximately 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% of the full-length protein. In another embodiment, the fragment comprises approximately 42%, 83%, 78%, or 66% of the full-length protein (excluding the signal sequence), as is described herein, in one embodiment, for HSV-2 gE, gC, gD, and gI.

[0061] In some embodiments, the fragment is a section of the protein, peptide, or nucleic acid. In another embodiment, the fragment is an immunogenic section of the protein, peptide or nucleic acid. In another embodiment, the fragment is a functional section within the protein, peptide or nucleic acid. In another embodiment, the fragment is an N-terminal immunogenic fragment. In some embodiments, the fragment is a C-terminal immunogenic fragment. In another embodiment, the fragment is an N-terminal functional fragment. In another embodiment, the fragment is a C-terminal functional fragment. In another embodiment, the fragment contains pieces of the protein linked together or pieces of multiple proteins linked together. In some embodiments, a fragment is a domain (e.g., an ectodomain).

[0062] As used herein, the term “functional” refers to the innate ability of a protein, peptide, nucleic acid, fragment or a variant thereof to exhibit a biological activity or function. In some embodiments, such a biological function is its binding property to an interaction partner, e.g., a membrane-associated receptor, and in another embodiment, its trimerization property. In the case of functional fragments and the functional variants of the disclosure, these biological functions may in fact be changed, e.g., with respect to their specificity or selectivity, but with retention of the basic biological function.

[0063] As used herein, the term “homology,” “homologous,” etc, when in reference to any protein or peptide, refer, in one embodiment, to a percentage of amino acid residues in the candidate sequence that are identical with the residues of a corresponding native polypeptide, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology, and not considering any conservative substitutions as part of the sequence identity. Methods and computer programs for the alignment are well known in the art.

[0064] In some embodiments, “homology” refers to identity of a protein sequence encoded by a modified (e.g., nucleoside-modified) polyribonucleotide to a sequence disclosed herein of greater than 70%. In another embodiment, the identity is greater than 72%. In another embodiment, the identity is greater than 75%. In another embodiment, the identity is greater than 78%. In another embodiment, the identity is greater than 80%. In another embodiment, the identity is greater than 82%. In another embodiment, the identity is greater than 83%. In another embodiment, the identity is greater than 85%. In another embodiment, the identity is greater than 87%. In another embodiment, the identity is greater than 88%. In another embodiment, the identity is greater than 90%. In another embodiment, the identity is greater than 92%. In another embodiment, the identity is greater than 93%. In another embodiment, the identity is greater than 95%. In another embodiment, the identity is greater than 96%. In another embodiment, the identity is greater than 97%. In another embodiment, the identity is greater than 98%. In another embodiment, the identity is greater than 99%. In another embodiment, the identity is 100%.

[0065] As used herein, the term “HSV-1” refers to a Herpes Simplex Virus-1. In some embodiments, “HSV-1” refers to a particular HSV-1 strain. In another embodiment, the term refers to a KOS strain. In another embodiment, the term refers to an F strain. In another embodiment, the term refers to an NS strain. In another embodiment, the term refers toPage 13 of 182 12197519v1a CL101 strain. In another embodiment, the term refers to a “17” strain. In another embodiment, the term refers to a “17+syn” strain. In another embodiment, the term refers to a MacIntyre strain. In another embodiment, the term refers to an MP strain. In another embodiment, the term refers to an HF strain. In another embodiment, the term refers to any other HSV-1 strain known in the art.

[0066] As used herein, the term “HSV-2” refers to a Herpes Simplex Virus-2. In some embodiments, “HSV-2” refers to a particular HSV-2 strain. In another embodiment, the term refers to an HSV-2333 strain. In another embodiment, the term refers to a 2.12 strain. In another embodiment, the term refers to an HG52 strain. In another embodiment, the term refers to an MS strain. In another embodiment, the term refers to a G strain. In another embodiment, the term refers to a 186 strain. In another embodiment, the term refers to any other HSV-2 strain known in the art.

[0067] As used herein, the term “humoral immunity” or “humoral immune response” refers to antibody production and the accessory processes that accompany it, including: Th2 activation and cytokine production, germinal center formation and isotype switching, affinity maturation and memory cell generation. It also refers to the effector functions of antibodies, which include pathogen neutralization, classical complement activation, and opsonin promotion of phagocytosis and pathogen elimination.

[0068] As used herein, the term “identity” refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules are considered to be “substantially identical” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical. Calculation of the percent identity of two nucleic acid or polypeptide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, 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 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of a reference sequence. The nucleotides at corresponding positions are then compared. When a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0). In some exemplary embodiments, nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix.Page 14 of 182 12197519v1

[0069] As used herein, the term “immune evasion domain” refers to a domain that interferes with or reduces in vivo anti-HSV efficacy of anti-HSV antibodies (e.g., anti-gE antibodies). In some embodiments, the domain interferes or reduces in vivo anti-HSV efficacy of an anti-HSV immune response. In some embodiments, the domain reduces the immunogenicity of an HSV protein (e.g., gE) during subsequent infection. In some embodiments, the domain reduces the immunogenicity of an HSV protein during subsequent challenge. In some embodiments, the domain reduces the immunogenicity of HSV during subsequent challenge. In some embodiments, the domain reduces the immunogenicity of an HSV protein in the context of ongoing HSV infection. In some embodiments, the domain reduces the immunogenicity of HSV in the context of ongoing HSV infection. In some embodiments, the domain functions as an IgG Fc receptor. In some embodiments, the domain promotes antibody bipolar bridging, which in one embodiment, is a term that refers to an antibody molecule binding by its Fab domain to an HSV antigen and by its Fc domain to a separate HSV antigen, such as in one embodiment, gE, thereby blocking the ability of the Fc domain to activate complement.

[0070] As used herein, the term “immunogenic fragment” refers to a portion of a protein that is in one embodiment immunogenic and in other embodiments elicits a protective immune response when administered to a subject.

[0071] As used herein, the term "immunogenicity" or "immunogenic" is used herein to refer to the innate ability of a protein, peptide, nucleic acid, antigen or organism to elicit an immune response in an animal when the protein, peptide, nucleic acid, antigen or organism is administered to the animal. In one embodiment, an immunogenic polypeptide is also antigenic. Thus, “enhancing the immunogenicity” in some embodiments, refers to increasing the ability of a protein, peptide, nucleic acid, antigen or organism to elicit an immune response in an animal when the protein, peptide, protein fragment, nucleic acid, antigen or organism is administered to an animal. The increased ability of a protein, peptide, protein fragment, nucleic acid, antigen or organism to elicit an immune response can be measured by, in some embodiments, a greater number of antibodies to a protein, peptide, protein fragment, nucleic acid, antigen or organism, a greater diversity of antibodies to an antigen or organism, a greater number of T-cells specific for a protein, peptide, protein fragment, nucleic acid, antigen or organism, a greater cytotoxic or helper T-cell response to a protein, peptide, nucleic acid, antigen or organism, and the like.

[0072]

[0073] As used herein, the term “immunologically equivalent” means that an immunologically equivalent molecule such as the immunologically equivalent amino acid sequence exhibits the same or essentially the same immunological properties and / or exerts the same or essentially the same immunological effects, e.g., with respect to the type of the immunological effect. In the context of the present disclosure, in some embodiments, the term “immunologically equivalent” is used with respect to the immunological effects or properties of antigens or antigen variants used for immunization. For example, an amino acid sequence is immunologically equivalent to a reference amino acid sequence if said amino acid sequence when exposed to the immune system of a subject induces an immune reaction having a specificity of reacting with the reference amino acid sequence.

[0074] In some embodiments, an antigen receptor is an antibody or B cell receptor which binds to an epitope in an antigen. In some embodiments, an antibody or B cell receptor binds to native epitopes of an antigen.Page 15 of 182 12197519v1

[0075] As used herein, these terms or grammatically comparable comparative terms, indicate values that are relative to a comparable reference measurement. For example, in some embodiments, an assessed value achieved with a provided pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) may be “increased” relative to that obtained with a comparable reference pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine). Alternatively or additionally, in some embodiments, an assessed value achieved in a subject may be “increased” relative to that obtained in the same subject under different conditions (e.g., prior to or after an event; or presence or absence of an event such as administration of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) as described herein, or in a different, comparable subject (e.g., in a comparable subject that differs from the subject of interest in prior exposure to a condition, e.g., absence of administration of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) as described herein.). In some embodiments, comparative terms refer to statistically relevant differences (e.g., that are of a prevalence and / or magnitude sufficient to achieve statistical relevance). Those skilled in the art will be aware, or will readily be able to determine, in a given context, a degree and / or prevalence of difference that is required or sufficient to achieve such statistical significance. In some embodiments, the term “reduced” or equivalent terms refers to a reduction in the level of an assessed value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or higher, as compared to a comparable reference. In some embodiments, the term “reduced” or equivalent terms refers to a complete or essentially complete inhibition, i.e., a reduction to zero or essentially to zero. In some embodiments, the term “increased” or “induced” refers to an increase in the level of an assessed value by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500%, or higher, as compared to a comparable reference.

[0076] As used herein, the term “ionizable” refers to a compound or group or atom that is charged at a certain pH. In the context of an ionizable amino lipid, such a lipid or a function group or atom thereof bears a positive charge at a certain pH. In some embodiments, an ionizable amino lipid is positively charged at an acidic pH. In some embodiments, an ionizable amino lipid is predominately neutral at physiological pH values, e.g., in some embodiments about 7.0-7.4, but becomes positively charged at lower pH values. In some embodiments, an ionizable amino lipid may have a pKa within a range of about 5 to about 7.

[0077] As used herein, the term “isolated” means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not “isolated”, but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated”. An isolated nucleic acid or protein can exist in substantially purified form.

[0078] As used herein, the term “isoform” refers to a version of a molecule, for example, a protein, with only slight differences to another isoform of the same protein. In some embodiments, isoforms may be produced from different but related genes, or in some embodiments, may arise from the same gene by alternative splicing. In some embodiments, isoforms are caused by single nucleotide polymorphisms.

[0079] As used herein, the term “RNA lipid nanoparticle” refers to a nanoparticle comprising at least one lipid and RNA molecule(s). In some embodiments, an RNA lipid nanoparticle comprises at least one ionizable amino lipid. In some embodiments, an RNA lipid nanoparticle comprises at least one ionizable amino lipid, at least one helper lipid, and at least one polymer-conjugated lipid (e.g., PEG-conjugated lipid). In various embodiments, RNA lipidPage 16 of 182 12197519v1nanoparticles as described herein can have an average size (e.g., Z-average) of about 100 nm to 1000 nm, or about 200 nm to 900 nm, or about 200 nm to 800 nm, or about 250 nm to about 700 nm. In some embodiments of the present disclosure, RNA lipid nanoparticles can have a particle size (e.g., Z-average) of about 30 nm to about 200 nm, or about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 80 nm to about 100 nm, about 90 nm to about 100 nm, about 70 to about 90 nm, about 80 nm to about 90 nm, or about 70 nm to about 80 nm. In some embodiments, an average size of lipid nanoparticles is determined by measuring the particle diameter. In some embodiments, RNA lipid nanoparticles may be prepared by mixing lipids with RNA molecules described herein.

[0080] As used herein, the term “lipidoid” refers to a lipid-like molecule. In some embodiments, a lipoid is an amphiphilic molecule with one or more lipid-like physical properties. In the context of the present disclosure, the term lipid is considered to encompass lipidoids.

[0081] As used herein, the term “nanoparticle” refers to a particle having an average size suitable for parenteral administration. In some embodiments, a nanoparticle has a longest dimension (e.g., a diameter) of less than 1,000 nanometers (nm). In some embodiments, a nanoparticle may be characterized by a longest dimension (e.g., a diameter) of less than 300 nm. In some embodiments, a nanoparticle may be characterized by a longest dimension (e.g., a diameter) of less than 100 nm. In many embodiments, a nanoparticle may be characterized by a longest dimension between about 1 nm and about 100 nm, or between about 1 µm and about 500 nm, or between about 1 nm and 1,000 nm. In many embodiments, a population of nanoparticles is characterized by an average size (e.g., longest dimension) that is below about 1,000 nm, about 500 nm, about 100 nm, about 50 nm, about 40 nm, about 30 nm, about 20 nm, or about 10 nm and often above about 1 nm. In many embodiments, a nanoparticle may be substantially spherical so that its longest dimension may be its diameter. In some embodiments, a nanoparticle has a diameter of less than 100 nm as defined by the National Institutes of Health.

[0082] As used herein, the term “naturally occurring” as used herein refers to an entity that can be found in nature. For example, a peptide or nucleic acid that is present in an organism (including viruses) and can be isolated from a source in nature and which has not been intentionally modified by man in the laboratory is naturally occurring.

[0083] As used herein, the term “neutralization” refers to an event in which binding agents such as antibodies bind to a biological active site of a virus such as a receptor binding protein, thereby inhibiting the parasitic infection of cells. In some embodiments, the term “neutralization” refers to an event in which binding agents eliminate or significantly reduce ability of infecting cells.

[0084] As used herein, the term “nucleic acid” refers to a polymer of at least 10 nucleotides or more. In some embodiments, a nucleic acid is or comprises DNA. In some embodiments, a nucleic acid is or comprises RNA. In some embodiments, a nucleic acid is or comprises peptide nucleic acid (PNA). In some embodiments, a nucleic acid is or comprises a single stranded nucleic acid. In some embodiments, a nucleic acid is or comprises a double- stranded nucleic acid. In some embodiments, a nucleic acid comprises both single and double-stranded portions. In some embodiments, a nucleic acid comprises a backbone that comprises one or more phosphodiester linkages. In some embodiments, a nucleic acid comprises a backbone that comprises both phosphodiester and non-Page 17 of 182 12197519v1phosphodiester linkages. For example, in some embodiments, a nucleic acid may comprise a backbone that comprises one or more phosphorothioate or 5'-N-phosphoramidite linkages and / or one or more peptide bonds, e.g., as in a “peptide nucleic acid”. In some embodiments, a nucleic acid comprises one or more, or all, natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, a nucleic acid comprises on or more, or all, non-natural residues. In some embodiments, a non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo- pyrimidine, 3 -methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5 -propynyl-cytidine, C5-methylcytidine, 2- aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 6-O-methylguanine, 2- thiocytidine, methylated bases, intercalated bases, and combinations thereof). In some embodiments, a non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared to those in natural residues. In some embodiments, a nucleic acid has a nucleotide sequence that encodes a functional gene product such as an RNA or polypeptide. In some embodiments, a nucleic acid has a nucleotide sequence that comprises one or more introns. In some embodiments, a nucleic acid may be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro, reproduction in a recombinant cell or system, or chemical synthesis. In some embodiments, a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 1 10, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides long.

[0085] As used herein, the term “nucleotide” refers to its art-recognized meaning. When a number of nucleotides is used as an indication of size, e.g., of a polynucleotide, a certain number of nucleotides refers to the number of nucleotides on a single strand, e.g., of a polynucleotide.

[0086] As used herein, the term “patient” refers to any organism who is suffering or at risk of a disease or disorder or condition. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, a patient is a human. In some embodiments, a patient is suffering from or susceptible to one or more diseases or disorders or conditions. In some embodiments, a patient displays one or more symptoms of a disease or disorder or condition. In some embodiments, a patient has been diagnosed with one or more diseases or disorders or conditions. In some embodiments, a disease or disorder or condition that is amenable to provided technologies is or includes an HSV infection. In some embodiments, a patient is receiving or has received certain therapy to diagnose and / or to treat a disease, disorder, or condition. In some embodiments, a patient is a patient suffering from or susceptible to an HSV infection.

[0087] As used herein, the term “pharmaceutical composition” refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments,Page 18 of 182 12197519v1pharmaceutical compositions may be specially formulated for parenteral administration, for example, by subcutaneous, intramuscular, or intravenous injection as, for example, a sterile solution or suspension formulation.

[0088] As used herein, the term “pharmaceutically effective amount” or “therapeutically effective amount” refers to the amount which achieves a desired reaction or a desired effect alone or together with further doses. In the case of the treatment of a particular disease, a desired reaction in some embodiments relates to inhibition of the course of the disease. In some embodiments, such inhibition may comprise slowing down the progress of a disease and / or interrupting or reversing the progress of the disease. In some embodiments, a desired reaction in a treatment of a disease may be or comprise delay or prevention of the onset of a disease or a condition. An effective amount of pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) described herein will depend, for example, on a disease or condition to be treated, the severity of such a disease or condition, individual parameters of the patient, including, e.g., age, physiological condition, size and weight, the duration of treatment, the type of an accompanying therapy (if present), the specific route of administration and similar factors. Accordingly, doses of pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) described herein may depend on various of such parameters. In the case that a reaction in a patient is insufficient with an initial dose, higher doses (or effectively higher doses achieved by a different, more localized route of administration) may be used.

[0089] As used herein, the term “poly(A) sequence” or “poly-A tail” refers to an uninterrupted or interrupted sequence of adenylate residues which is typically located at the 3'-end of an RNA molecule. Poly(A) sequences are known to those of skill in the art and may follow the 3’-UTR in the RNAs described herein. An uninterrupted poly(A) sequence is characterized by consecutive adenylate residues. In nature, an uninterrupted poly(A) sequence is typical. RNAs disclosed herein can have a poly(A) sequence attached to the free 3'-end of the RNA by a template- independent RNA polymerase after transcription or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase.

[0090] As used herein, the term “prevent” or “prevention” when used in connection with the occurrence of a disease, disorder, and / or condition, refers to reducing the risk of developing the disease, disorder and / or condition and / or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time.

[0091] As used herein, the term “properdin-interfering domain” refers to a domain that blocks or inhibits binding of a host C3b molecule with a host properdin molecule. In another embodiment, the term refers to a domain that blocks or inhibits an interaction of a host C3b molecule with a host properdin molecule.

[0092] As used herein the term “recombinant” in the context of the present disclosure means “made through genetic engineering”. In some embodiments, a “recombinant” entity such as a recombinant nucleic acid in the context of the present disclosure is not naturally occurring.

[0093] As used herein, the term “reference” describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and / or determined substantially simultaneously with thePage 19 of 182 12197519v1testing or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and / or comparison to a particular possible reference or control.

[0094] As will be understood from context, “risk” of a disease, disorder, and / or condition refers to a likelihood that a particular individual will develop the disease, disorder, and / or condition. In some embodiments, risk is expressed as a percentage. In some embodiments, risk is from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90 up to 100%. In some embodiments risk is expressed as a risk relative to a risk associated with a reference sample or group of reference samples. In some embodiments, a reference sample or group of reference samples have a known risk of a disease, disorder, condition and / or event. In some embodiments a reference sample or group of reference samples are from individuals comparable to a particular individual. In some embodiments, relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. In some embodiments, risk may reflect one or more genetic attributes, e.g., which may predispose an individual toward development (or not) of a particular disease, disorder and / or condition. In some embodiments, risk may reflect one or more epigenetic events or attributes and / or one or more lifestyle or environmental events or attributes.

[0095] The term “selective” or “specific”, when used herein in reference to an agent having an activity, is understood by those skilled in the art to mean that the agent discriminates between potential target entities, states, or cells. For example, in some embodiments, an agent is said to bind “specifically” to its target if it binds preferentially with that target in the presence of one or more competing alternative targets. In many embodiments, specific interaction is dependent upon the presence of a particular structural feature of the target entity (e.g., an epitope, a cleft, a binding site). It is to be understood that specificity need not be absolute. In some embodiments, specificity may be evaluated relative to that of a target-binding moiety for one or more other potential target entities (e.g., competitors). In some embodiments, specificity is evaluated relative to that of a reference specific binding moiety. In some embodiments, specificity is evaluated relative to that of a reference non-specific binding moiety.

[0096] As used herein, the term “subject” refers to an organism to be administered with a composition described herein, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans. In some embodiments, a subject is a human subject. In some embodiments, a subject is suffering from a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject is susceptible to a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject displays one or more non- specific symptoms of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject is a patient. In some embodiments, a subject is an individual to whom diagnosis and / or therapy is and / or has been administered.Page 20 of 182 12197519v1

[0097] As used herein, an individual who is “suffering from” a disease, disorder, and / or condition has been diagnosed with and / or displays one or more symptoms of a disease, disorder, and / or condition.

[0098] As used herein, an individual who is “susceptible to” a disease, disorder, and / or condition is one who has a higher risk of developing the disease, disorder, and / or condition than does a member of the general public. In some embodiments, an individual who is susceptible to a disease, disorder and / or condition may not have been diagnosed with the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will develop the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will not develop the disease, disorder, and / or condition.

[0099] As used herein, the term “synthetic” refers to an entity that is artificial, or that is made with human intervention, or that results from synthesis rather than naturally occurring. For example, in some embodiments, a synthetic nucleic acid or polynucleotide refers to a nucleic acid molecule that is chemically synthesized, e.g., in some embodiments by solid-phase synthesis. In some embodiments, the term “synthetic” refers to an entity that is made outside of biological cells. For example, in some embodiments, a synthetic nucleic acid or polynucleotide refers to a nucleic acid molecule (e.g., an RNA) that is produced by in vitro transcription using a template.

[0100] As used herein, the term “therapy” refers to an administration or delivery of an agent or intervention that has a therapeutic effect and / or elicits a desired biological and / or pharmacological effect (e.g., has been demonstrated to be statistically likely to have such effect when administered to a relevant population). In some embodiments, a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. In some embodiments, a therapeutic agent or therapy is a medical intervention (e.g., surgery, radiation, phototherapy) that can be performed to alleviate, relieve, inhibit, present, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition.

[0101] As used herein, the terms “three prime untranslated region” or “3' UTR” refer to a sequence of an RNA molecule that begins following a stop codon of a coding region of an open reading frame sequence. In some embodiments, the 3' UTR begins immediately after a stop codon of a coding region of an open reading frame sequence, e.g., in its natural context. In other embodiments, the 3' UTR does not begin immediately after stop codon of the coding region of an open reading frame sequence, e.g., in its natural context.

[0102] As used herein, the term “threshold level” refers to a level that are used as a reference to attain information on and / or classify the results of a measurement, for example, the results of a measurement attained in an assay. For example, in some embodiments, a threshold level means a value measured in an assay that defines the dividing line between two subsets of a population (e.g. a batch that satisfy quality control criteria vs. a batch that does not satisfy quality control criteria). Thus, a value that is equal to or higher than the threshold level defines one subset of the population, and a value that is lower than the threshold level defines the other subset of the population. A threshold level can be determined based on one or more control samples or across a population ofPage 21 of 182 12197519v1control samples. A threshold level can be determined prior to, concurrently with, or after the measurement of interest is taken. In some embodiments, a threshold level can be a range of values.

[0103] As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and / or condition, for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject at a later-stage of disease, disorder, and / or condition.

[0104] As used herein, the term “truncated” refers to a protein or polypeptide that is shorter at its N-terminal, C- terminal, or both the N-terminal and C-terminal ends as compared to the wild-type protein or polypeptide.

[0105] As used herein, the term “vaccination” refers to the administration of a composition intended to generate an immune response, for example to a disease-associated (e.g., disease-causing) agent. In some embodiments, vaccination can be administered before, during, and / or after exposure to a disease-associated agent, and in certain embodiments, before, during, and / or shortly after exposure to the agent. In some embodiments, vaccination includes multiple administrations, appropriately spaced in time, of a vaccine composition. In some embodiments, vaccination generates an immune response to an infectious agent.

[0106] As used herein, the term “vaccine” refers to a composition or a combination or an agent that upon administration to an individual stimulates antibody production or cellular immunity against a pathogen but is incapable of causing severe infection. In some embodiments, the vaccine is incapable of causing severe infection in a subject.

[0107] As used herein in the context of molecules, e.g., nucleic acids, proteins, or small molecules, the term “variant” refers to a molecule that shows significant structural identity with a reference molecule but differs structurally from the reference molecule, e.g., in the presence or absence or in the level of one or more chemical moieties as compared to the reference entity. In some embodiments, a variant also differs functionally from its reference molecule. In general, whether a particular molecule is properly considered to be a “variant” of a reference molecule is based on its degree of structural identity with the reference molecule. As will be appreciated by those skilled in the art, any biological or chemical reference molecule has certain characteristic structural elements. A variant, by definition, is a distinct molecule that shares one or more such characteristic structural elements but differs in at least one aspect from the reference molecule. In some embodiments, a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and / or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nucleic acid (e.g., that are attached to the polypeptide or nucleic acid backbone). In some embodiments, a variant polypeptide or nucleic acid shows an overall sequence identity with a reference polypeptide or nucleic acid that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 99%. In some embodiments, a variant polypeptide or nucleic acid does not share at least one characteristic sequence element with a reference polypeptide or nucleic acid. In some embodiments, a referencePage 22 of 182 12197519v1polypeptide or nucleic acid has one or more biological activities. In some embodiments, a variant polypeptide or nucleic acid shares one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid lacks one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid shows a reduced level of one or more biological activities as compared to the reference polypeptide or nucleic acid. In some embodiments, a polypeptide or nucleic acid of interest is considered to be a “variant” of a reference polypeptide or nucleic acid if it has an amino acid or nucleotide sequence that is identical to that of the reference but for a small number of sequence alterations at particular positions. Typically, fewer than about 20%, about 15%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, or about 2% of the residues in a variant are substituted, inserted, or deleted, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1 substituted residue as compared to a reference. Often, a variant polypeptide or nucleic acid comprises a very small number (e.g., fewer than about 5, about 4, about 3, about 2, or about 1) number of substituted, inserted, or deleted, functional residues (i.e., residues that participate in a particular biological activity) relative to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises not more than about 5, about 4, about 3, about 2, or about 1 addition or deletion, and, in some embodiments, comprises no additions or deletions, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises fewer than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and commonly fewer than about 5, about 4, about 3, or about 2 additions or deletions as compared to the reference. In some embodiments, a reference polypeptide or nucleic acid is one found in nature.

[0108] In some embodiments, the variant may be a sequence conservative variant. In some embodiments, the variant may be a functional conservative variant. In some embodiments, a variant may comprise an addition, deletion or substitution of one or more amino acids.

[0109] As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid”, which refers to a circular double stranded DNA loop into which additional DNA segments may be ligated. Another type of vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as “expression vectors.” In some embodiments, known techniques may be used, for example, for generation or manipulation of recombinant DNA, for oligonucleotide synthesis, and for tissue culture and transformation (e.g., electroporation, lipofection). Enzymatic reactions and purification techniques may be performed according to manufacturer's specifications or as commonly accomplished in the art or as described herein. The foregoing techniques and procedures may be generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification. See e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual (4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2012)), which is incorporated herein by reference for any purpose.Page 23 of 182 12197519v1

[0110] All literature and similar material cited in this application, including, but not limited to, patents, patent applications, articles, books, treatises, and web pages, regardless of the format of such literature and similar materials, are expressly incorporated by reference in their entirety. In the event that one or more of the incorporated literature and similar materials differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, this application controls. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described in any way. Polyribonucleotides Encoding HSV Polypeptides

[0111] In some embodiments, the present disclosure provides a polyribonucleotide encoding a polypeptide that comprises a viral antigen, or an immunogenic fragment thereof, from Herpes Simplex Virus (HSV). In some embodiments, the present disclosure provides a nucleoside-modified polyribonucleotide encoding a polypeptide that comprises a viral antigen, or an immunogenic fragment thereof, from Herpes Simplex Virus (HSV). In some embodiments, the term “polyribonucleotide” as used herein can be used interchangeably with “RNA”. In some embodiments, the HSV viral antigen, or immunogenic fragment thereof, is from HSV-2. In another embodiment, the HSV viral antigen, or immunogenic fragment thereof, is from HSV-1. Thus, in some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding a polypeptide that comprises a viral antigen, or an immunogenic fragment thereof, from Herpes Simplex Virus-2 (HSV-2). HSV Glycoprotein B

[0112] In some embodiments, the present disclosure provides a polyribonucleotide or RNA encoding a polypeptide that comprises glycoprotein B (gB) of HSV, or an immunogenic fragment thereof. In some embodiments, the polyribonucleotide or RNA encodes a polypeptide that comprises gB of HSV-1 (gB1), or an immunogenic fragment thereof. In another embodiment, the polyribonucleotide or RNA encodes a polypeptide that comprises gB of HSV-2 (gB2), or an immunogenic fragment thereof.

[0113] In some embodiments, the present disclosure provides a nucleoside-modified polyribonucleotide or RNA encoding a polypeptide that comprises glycoprotein B (gB) of HSV, or an immunogenic fragment thereof. In some embodiments, the nucleoside-modified polyribonucleotide or RNA encodes a polypeptide that comprises gB of HSV-1 (gB1), or an immunogenic fragment thereof. In another embodiment, the nucleoside-modified polyribonucleotide or RNA encodes a polypeptide that comprises gB of HSV-2 (gB2), or an immunogenic fragment thereof.

[0114] In some embodiments, the present disclosure provides an RNA encoding a polypeptide that comprises full length HSV-1 gB. In some embodiments, the present disclosure provides a nucleoside-modified RNA encoding a polypeptide that comprises full length HSV-1 gB. In some embodiments, the amino acid sequence of full length HSV- 1 gB comprises the sequence as set forth in SEQ ID NO: 8 or SEQ ID NO: 11. In some embodiments, an HSV-1 gB fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 8 or SEQ ID NO: 11. In another embodiment, the amino acid sequence of full length HSV-1 gB sequence comprises the sequence as set forth in SEQ ID NO: 9. In some embodiments, thePage 24 of 182 12197519v1nucleotide sequence of HSV-1 gB comprises a nucleotide sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 9.

[0115] In another embodiment, the amino acid sequence of an HSV-1 gB antigen sequence comprises the sequence as set forth in SEQ ID NO: 199. In some embodiments, amino acid sequence of an HSV-1 gB antigen is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 199. In another embodiment, the amino acid sequence of an HSV-1 gB antigen sequence comprises the sequence as set forth in SEQ ID NO: 200. In some embodiments, amino acid sequence of an HSV-1 gB antigen is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 200.

[0116] In some embodiments, the present disclosure provides an RNA encoding a polypeptide that comprises full length HSV-2 gB. In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding a polypeptide that comprises full length HSV-2 gB. In some embodiments, the amino acid sequence of full length HSV-2 gB comprises the sequence as set forth in SEQ ID NO: 7. In some embodiments, an HSV-1 gB fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 7.

[0117] In another embodiment, the amino acid sequence of an HSV-2 gB antigen sequence comprises the sequence as set forth in SEQ ID NO: 201. In some embodiments, amino acid sequence of an HSV-2 gB antigen is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 201. In another embodiment, the amino acid sequence of an HSV-2 gB antigen sequence comprises the sequence as set forth in SEQ ID NO: 202. In some embodiments, amino acid sequence of an HSV-2 gB antigen is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 202.

[0118] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding a polypeptide that comprises the ectodomain of HSV-1 gB, or an immunogenic fragment thereof. In some embodiments, the ectodomain of HSV-1 gB is as set forth in SEQ ID NO: 10.

[0119] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 203. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 203.

[0120] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding atPage 25 of 182 12197519v1least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 204. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 204.

[0121] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 205. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 205.

[0122] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 206. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 206.

[0123] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 207. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 207.

[0124] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 208. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 208.

[0125] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 209. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 209.

[0126] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 210. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 210.

[0127] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 211. In some embodiments, an RNA or a nucleoside-modified RNA encoding at leastPage 26 of 182 12197519v1an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 211.

[0128] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 212. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 212.

[0129] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 213. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 213.

[0130] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 214. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 214.

[0131] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 215. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 215.

[0132] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 216. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 216.

[0133] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 217. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 217.

[0134] In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB comprises SEQ ID NO: 218. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-1 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 218.

[0135] In another embodiment, the present disclosure provides an RNA or a nucleoside-modified RNA encoding a polypeptide that comprises the ectodomain of HSV-2 gB, or an immunogenic fragment thereof. In some embodiments, the ectodomain of HSV-2 gB comprises the amino acid sequence as set forth in SEQ ID NO: 1. In some embodiments, the ectodomain of HSV-2 gB consists of or consists essentially of the amino acid sequence of SEQ ID NO: 1. In some embodiments, the present disclosure encompasses sequences that are at least 80% (e.g., at least 85%, 90%, 95%, 96%, 97%, 98%, or 99%) identical to SEQ ID NO: 1. In some embodiments, the ectodomain of HSV-2 gB is encoded by a nucleotide sequence comprising the sequence of SEQ ID NO: 5. In some embodiments,Page 27 of 182 12197519v1the ectodomain of HSV-2 gB is encoded by a nucleotide sequence consisting of or consisting essentially of SEQ ID NO: 5. In some embodiments, the present disclosure encompasses sequences that are at least 80% (e.g., at least 85%, 90%, 95%, 96%, 97%, 98%, or 99%) identical to SEQ ID NO: 5.

[0136] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 219. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 219.

[0137] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 220. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 220.

[0138] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 221. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 221.

[0139] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 222. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 222.

[0140] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 223. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 223.

[0141] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 224. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, atPage 28 of 182 12197519v1least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 224.

[0142] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 225. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 225.

[0143] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 226. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 226.

[0144] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 227. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 227.

[0145] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 228. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 228.

[0146] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 229. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 229.

[0147] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB. In some embodiments, an RNA or a nucleoside-modified RNA encoding at least an immunogenic part of HSV-2 gB comprises SEQ ID NO: 230. In some embodiments, an RNA or a nucleoside- modified RNA encoding at least an immunogenic part of HSV-2 gB is at least 85%, at least 90%, at least 91%, atPage 29 of 182 12197519v1least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 230. Additional HSV Glycoproteins

[0148] In some embodiments, in addition to an RNA or a nucleoside-modified RNA encoding an HSV gB, or immunogenic fragment thereof, compositions of the present disclosure may further comprise one or more RNAs

[0149] In some embodiments, compositions of the present disclosure further comprise an RNA or a nucleoside- modified RNA encoding a polypeptide comprising, or consisting of, HSV glycoprotein C (gC), or an immunogenic fragment thereof.

[0150] In some embodiments, disclosed herein is a single composition comprising an RNA encoding an HSV gB and an RNA encoding an HSV gC, or immunogenic fragments thereof. In some embodiments, said gB and said gC RNAs are part of a single polyribonucleotide. In some embodiments, disclosed herein are two individual compositions to be administered as part of a single treatment, the first composition comprising an RNA encoding an HSV gB or an immunogenic fragment thereof, and the second composition comprising an RNA encoding an HSV gC or immunogenic fragment thereof. In some embodiments, an RNA encoding an HSV gC, or an immunogenic fragment thereof, comprises an RNA encoding HSV-1 gC, also termed gC1, or an immunogenic fragment thereof. In some embodiments, a nucleotide sequence of the RNA encoding an HSV-1 gC fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAA UCAUUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGGCCAUCUCCGGCGUGCCC GUGCUGGGCUUCUUCAUCAUCGCCGUGCUGAUGUCCGCCCAGGAGUCCUGGGCCGAGACCGCCUCCACCGGCCCC ACCAUCACCGCCGGCGCCGUGACCAACGCCUCCGAGGCCCCCACCUCCGGCUCCCCCGGCUCCGCCGCCUCCCCCGAGGUGAC CCCCACCUCCACCCCCAACCCCAACAACGUGACCCAGAACAAGACCACCCCCACCGAGCCCGCCUCCCCCCCCACCACCCCCAAG CCCACCUCCACCCCCAAGUCCCCCCCCACCUCCACCCCCGACCCCAAGCCCAAGAACAACACCACCCCCGCCAAGUCCGGCCGCC CCACCAAGCCCCCCGGCCCCGUGUGGUGCGACCGCCGCGACCCCCUGGCCCGCUACGGCUCCCGCGUGCAGAUCCGCUGCCGC UUCCGCAACUCCACCCGCAUGGAGUUCCGCCUGCAGAUCUGGCGCUACUCCAUGGGCCCCUCCCCCCCCAUCGCCCCCGCCCC CGACCUGGAGGAGGUGCUGACCAACAUCACCGCCCCCCCCGGCGGCCUGCUGGUGUACGACUCCGCCCCCAACCUGACCGACC CCCACGUGCUGUGGGCCGAGGGCGCCGGCCCCGGCGCCGACCCCCCCCUGUACUCCGUGACCGGCCCCCUGCCCACCCAGCGC CUGAUCAUCGGCGAGGUGACCCCCGCCACCCAGGGCAUGUACUACCUGGCCUGGGGCCGCAUGGACUCCCCCCACGAGUACG GCACCUGGGUGCGCGUGCGCAUGUUCCGCCCCCCCUCCCUGACCCUGCAGCCCCACGCCGUGAUGGAGGGCCAGCCCUUCAA GGCCACCUGCACCGCCGCCGCCUACUACCCCCGCAACCCCGUGGAGUUCGACUGGUUCGAGGACGACCGCCAGGUGUUCAACC CCGGCCAGAUCGACACCCAGACCCACGAGCACCCCGACGGCUUCACCACCGUGUCCACCGUGACCUCCGAGGCCGUGGGCGGC CAGGUGCCCCCCCGCACCUUCACCUGCCAGAUGACCUGGCACCGCGACUCCGUGACCUUCUCCCGCCGCAACGCCACCGGCCU GGCCCUGGUGCUGCCCCGCCCCACCAUCACCAUGGAGUUCGGCGUGCGCCACGUGGUGUGCACCGCCGGCUGCGUGCCCGAG GGCGUGACCUUCGCCUGGUUCCUGGGCGACGACCCCUCCCCCGCCGCCAAGUCCGCCGUGACCGCCCAGGAGUCCUGCGACCPage 30 of 182 12197519v1ACCCCGGCCUGGCCACCGUGCGCUCCACCCUGCCCAUCUCCUACGACUACUCCGAGUACAUCUGCCGCCUGACCGGCUACCCC GCCGGCAUCCCCGUGCUGGAGCACCACUAACUAGUAGUGACUGACUAGGAUCUGGUUACCACUAAACCAGCCUCAAGAACACC CGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCC UAAUAAAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 12)

[0151] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 13). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 14). In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 15) and poly adenylation tail (SEQ ID NO: 16).

[0152] In some embodiments, the polyribonucleotide encoding an HSV-1 gC signal sequence MHCII comprises the sequence ATGGCCATCTCCGGCGTGCCCGTGCTGGGCTTCTTCATCATCGCCGTGCTGATGTCCGCCCAGGAGTCCTGGGCC (SEQ ID NO: 14).

[0153] In some embodiments, a 3’ UTR comprises the sequence CTAGTAGTGACTGACTAGGATCTGGTTACCACTAAACCAGCCTCAAGAACACCCGAATGGAGTCTCTAAGCTACATAATACCAAC TTACACTTACAAAATGTTGTCCCCCAAAATGTAGCCATTCGTATCTGCTCCTAATAAAAAGAAAGTTTCTTCACATTCT (SEQ ID NO: 15).

[0154] In another embodiment, the nucleotide sequence of the RNA encoding an HSV-1 gC fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof. In some embodiments, the sequence of the HSV-1 gC fragment is as set forth in SEQ ID NO: 17.

[0155] In some embodiments, the HSV-1 gC fragment encoded by an RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-457 of gC from HSV-1 (e.g., KOS strain), as set forth in the following amino acid sequence: ETASTGPTITAGAVTNASEAPTSGSPGSAASPEVTPTSTPNPNNVTQNKTTPTEPASPPTTPKPTSTPKSPPTSTPDPKPKNNTTPAK SGRPTKPPGPVWCDRRDPLARYGSRVQIRCRFRNSTRMEFRLQIWRYSMGPSPPIAPAPDLEEVLTNITAPPGGLLVYDSAPNLTDP HVLWAEGAGPGADPPLYSVTGPLPTQRLIIGEVTPATQGMYYLAWGRMDSPHEYGTWVRVRMFRPPSLTLQPHAVMEGQPFKATC TAAAYYPRNPVEFDWFEDDRQVFNPGQIDTQTHEHPDGFTTVSTVTSEAVGGQVPPRTFTCQMTWHRDSVTFSRRNATGLALVLP RPTITMEFGVRHVVCTAGCVPEGVTFAWFLGDDPSPAAKSAVTAQESCDHPGLATVRSTLPISYDYSEYICRLTGYPAGIPVLEHH (SEQ ID NO: 18).

[0156] In some embodiments, an HSV-1 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 18. In some embodiments, an HSV-1 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 18. In some embodiments, the gC fragment encoded by an RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-457 of gC from an HSV-1 strain.Page 31 of 182 12197519v1

[0157] In some embodiments, the HSV-1 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 25-457 of gC from HSV-1 (e.g., KOS strain), as set forth in the following amino acid sequence: GSETASTGPTITAGAVTNASEAPTSGSPGSAASPEVTPTSTPNPNNVTQNKTTPTEPASPPTTPKPTSTPKSPPTSTPDPKPKNNTTP AKSGRPTKPPGPVWCDRRDPLARYGSRVQIRCRFRNSTRMEFRLQIWRYSMGPSPPIAPAPDLEEVLTNITAPPGGLLVYDSAPNLT DPHVLWAEGAGPGADPPLYSVTGPLPTQRLIIGEVTPATQGMYYLAWGRMDSPHEYGTWVRVRMFRPPSLTLQPHAVMEGQPFKA TCTAAAYYPRNPVEFDWFEDDRQVFNPGQIDTQTHEHPDGFTTVSTVTSEAVGGQVPPRTFTCQMTWHRDSVTFSRRNATGLALV LPRPTITMEFGVRHVVCTAGCVPEGVTFAWFLGDDPSPAAKSAVTAQESCDHPGLATVRSTLPISYDYSEYICRLTGYPAGIPVLEHH (SEQ ID NO: 19).

[0158] In some embodiments, an HSV-1 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 19. In some embodiments, an HSV-1 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 19.

[0159] In some embodiments, the full-length HSV-1 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MAPGRVGLAVVLWGLLWLGAGVAGGSETASTGPTITAGAVTNASEAPTSGSPGSAASPEVTPTSTPNPNNVTQNKTTPTEPASPPT TPKPTSTPKSPPTSTPDPKPKNNTTPAKSGRPTKPPGPVWCDRRDPLARYGSRVQIRCRFRNSTRMEFRLQIWRYSMGPSPPIAPAP DLEEVLTNITAPPGGLLVYDSAPNLTDPHVLWAEGAGPGADPPLYSVTGPLPTQRLIIGEVTPATQGMYYLAWGRMDSPHEYGTWV RVRMFRPPSLTLQPHAVMEGQPFKATCTAAAYYPRNPVEFDWFEDDRQVFNPGQIDTQTHEHPDGFTTVSTVTSEAVGGQVPPRT FTCQMTWHRDSVTFSRRNATGLALVLPRPTITMEFGVRHVVCTAGCVPEGVTFAWFLGDDPSPAAKSAVTAQESCDHPGLATVRST LPISYDYSEYICRLTGYPAGIPVLEHHGSHQPPPRDPTERQVIEAIEWVGIGIGVLAAGVLVVTAIVYVVRTSQSRQRHRR (SEQ ID NO: 20).

[0160] In some embodiments, an HSV-1 gC comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 20. In some embodiments, an HSV-1 gC has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 20.

[0161] In other embodiments, the HSV-1 gC, or an immunogenic fragment thereof, encoded by an RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA45779.1, AAA96680.1, ABI63505.1, ABM52973.1, ABM52976.1, ABM52977.1, ACM62267.1, ADD60042.1, ADD60119.1, ADM22367.1, ADM22444.1, ADM22520.1, ADM22597.1, ADM22674.1, ADM22751.1, ADM22827.1, ADM22904.1, ADM22981.1, ADM23057.1, ADM23133.1, ADM23210.1, ADM23287.1, ADM23361.1, ADM23435.1, ADM23509.1, ADM23583.1, ADM23658.1, ADM23733.1, ADM23809.1, AEQ77075.1, AEQ77099.1, AER37628.1, AER37697.1, AER37767.1, AER37838.1, AER37910.1, AER37981.1, AER38051.2, AFA36179.1, AFA36180.1, AFA36181.1, AFA36182.1, AFA36183.1, AFA36184.1, AFA36185.1, AFA36186.1, AFA36187.1, AFA36188.1, AFA36189.1, AFA36190.1, AFA36191.1, AFA36192.1, AFA36193.1, AFA36194.1, AFA36195.1, AFA36196.1, AFA36197.1, AFA36198.1, AFA36199.1, AFA36200.1, AFA36201.1, AFA36202.1, AFA36203.1, AFE62872.1, AFH78104.1, AFI23635.1, AFK50391.1, AFP86408.1, AGZ01906.1,Page 32 of 182 12197519v1AIR95840.1, AJE59989.1, AJE60060.1, AJE60131.1, AJE60202.1, AKE48623.1, AKE98415.1, AKE98416.1, AKE98417.1, AKE98418.1, AKE98419.1, AKE98420.1, AKE98421.1, AKE98422.1, AKE98423.1, AKE98424.1, AKE98425.1, AKE98426.1, AKE98427.1, AKE98428.1, AKE98429.1, AKE98430.1, AKE98431.1, AKE98432.1, AKE98433.1, AKE98434.1, AKE98435.1, AKG59227.1, AKG59299.1, AKG59372.1, AKG59444.1, AKG59516.1, AKG59591.1, AKG59663.1, AKG59736.1, AKG59807.1, AKG59879.1, AKG59953.1, AKG60027.1, AKG60099.1, AKG60170.1, AKG60243.1, AKG60316.1, AKG60386.1, AKG60456.1, AKG60528.1, AKG60601.1, AKG60674.1, AKG60745.1, AKG60817.1, AKG60887.1, AKG60959.1, AKG61032.1, AKG61104.1, AKG61175.1, AKG61248.1, AKG61321.1, AKG61392.1, AKG61464.1, AKG61537.1, AKG61611.1, AKG61684.1, AKG61756.1, AKG61828.1, AKG61902.1, AKG61974.1, AKH80444.1, AKH80517.1, AKM76368.1, ALM22613.1, ALM22687.1, ALM22761.1, ALM22835.1, ALO18641.1, ALO18717.1, AMB65642.1, AMB65715.1, AMB65862.1, AMN09813.1, ANN83942.1, ANN84019.1, ANN84095.1, ANN84172.1, ANN84249.1, ANN84326.1, ANN84403.1, ANN84478.1, ANN84555.1, ANN84632.1, ANN84708.1, ANN84785.1, ANN84861.1, ANN84938.1, ANN85014.1, ANN85091.1, ANN85167.1, ANN85242.1, ANN85319.1, ANN85396.1, ANN85472.1, ANN85549.1, ANN85626.1, ANN85703.1, ANN85779.1, AOY34308.1, AOY36663.1, AOY36687.1, ARB08935.1, ARO38059.1, ARO38060.1, ARO38061.1, ARO38062.1, ARO38063.1, ARO38064.1, ARO38065.1, ARO38066.1, ASM47642.1, ASM47719.1, ASM47796.1, ASM47871.1, BAM73394.1, CAA32294.1, CAB40083.1, CAD13356.1, CAD13357.1, CAD13358.1, CAD13359.1, CAD13360.1, CAD13361.1, CAD13362.1, CAD13363.1, CAD13364.1, CAD13365.1, CAD13366.1, CAD13367.1, CAD13368.1, CAD13369.1, CAD13370.1, CAD13371.1, CAD13372.1, CAD13373.1, CAD13374.1, CAD13375.1, CAD13376.1, CAD13377.1, CAD13378.1, P04290.1, P04488.1, P09855.1, P10228.1, P28986.1, SBO07729.1, SBO07793.1, SBO07798.1, SBO07812.1, SBO07880.1, SBS69375.1, SBS69379.1, SBS69440.1, SBS69448.1, SBS69560.1, SBS69599.1, SBS69602.1, SBS69637.1, SBS69790.1, SBT69374.1, SCL76887.1, YP_009137119.1, or YP_009137143.1.

[0162] In some embodiments, an RNA encoding HSV gC as described herein comprises an RNA encoding HSV-2 gC, also termed gC2, or an immunogenic fragment thereof.

[0163] In some embodiments, the nucleotide sequence of an RNA encoding an HSV-2 gC fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAA UCAUUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCUGCUGCUG CUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCGCCUCCCCCGGCCGCACCAUCACCGUGGGCCCCCGCGGCAAC GCCUCCAACGCCGCCCCCUCCGCCUCCCCCCGCAACGCCUCCGCCCCCCGCACCACCCCCACCCCCCCCCAGCCCCGCAAGGCC ACCAAGUCCAAGGCCUCCACCGCCAAGCCCGCCCCCCCCCCCAAGACCGGCCCCCCCAAGACCUCCUCCGAGCCCGUGCGCUGC AACCGCCACGACCCCCUGGCCCGCUACGGCUCCCGCGUGCAGAUCCGCUGCCGCUUCCCCAACUCCACCCGCACCGAGUUCCG CCUGCAGAUCUGGCGCUACGCCACCGCCACCGACGCCGAGAUCGGCACCGCCCCCUCCCUGGAGGAGGUGAUGGUGAACGUG UCCGCCCCCCCCGGCGGCCAGCUGGUGUACGACUCCGCCCCCAACCGCACCGACCCCCACGUGAUCUGGGCCGAGGGCGCCGG CCCCGGCGCCUCCCCCCGCCUGUACUCCGUGGUGGGCCCCCUGGGCCGCCAGCGCCUGAUCAUCGAGGAGCUGACCCUGGAG ACCCAGGGCAUGUACUACUGGGUGUGGGGCCGCACCGACCGCCCCUCCGCCUACGGCACCUGGGUGCGCGUGCGCGUGUUCC GCCCCCCCUCCCUGACCAUCCACCCCCACGCCGUGCUGGAGGGCCAGCCCUUCAAGGCCACCUGCACCGCCGCCACCUACUAC CCCGGCAACCGCGCCGAGUUCGUGUGGUUCGAGGACGGCCGCCGCGUGUUCGACCCCGCCCAGAUCCACACCCAGACCCAGG AGAACCCCGACGGCUUCUCCACCGUGUCCACCGUGACCUCCGCCGCCGUGGGCGGCCAGGGCCCCCCCCGCACCUUCACCUGCPage 33 of 182 12197519v1CAGCUGACCUGGCACCGCGACUCCGUGUCCUUCUCCCGCCGCAACGCCUCCGGCACCGCCUCCGUGCUGCCCCGCCCCACCAU CACCAUGGAGUUCACCGGCGACCACGCCGUGUGCACCGCCGGCUGCGUGCCCGAGGGCGUGACCUUCGCCUGGUUCCUGGGC GACGACUCCUCCCCCGCCGAGAAGGUGGCCGUGGCCUCCCAGACCUCCUGCGGCCGCCCCGGCACCGCCACCAUCCGCUCCAC CCUGCCCGUGUCCUACGAGCAGACCGAGUACAUCUGCCGCCUGGCCGGCUACCCCGACGGCAUCCCCGUGCUGGAGCACCAC UAACUAGUAGUGACUGACUAGGAUCUGGUUACCACUAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUA CCAACUUACACUUACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAUUC UAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAA (SEQ ID NO: 21).

[0164] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 13). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 3). In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 15) and poly adenylation tail (SEQ ID NO: 16).

[0165] In another embodiment, the nucleotide sequence of the RNA encoding an HSV-2 gC fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof. In some embodiments, the sequence of the HSV-2 gC fragment is as set forth in SEQ ID NO: 22.

[0166] In some embodiments, the HSV-2 gC fragment encoded by an RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-426 of gC from HSV-2 (e.g., strain 333 or UL44), as set forth in the following amino acid sequence: ASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTGPPKTSSEPVRCNRHDPLARYGSRVQIRCR FPNSTRTESRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLIIE ELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYYPGNRAEFVWFEDGRRVFDPAQIHTQ TQENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWFLG DDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHH (SEQ ID NO: 23).

[0167] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 23. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 23.

[0168] In some embodiments, an HSV-2 gC fragment comprises the following amino acid sequence: ASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTGPPKTSSEPVRCNRHDPLARYGSRVQIRCR FPNSTRTEFRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLIIE ELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYYPGNRAEFVWFEDGRRVFDPAQIHTQ TQENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWFLG DDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHH (SEQ ID NO: 24).

[0169] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at leastPage 34 of 182 12197519v198%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 24. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 24.

[0170] In some embodiments, the HSV-2 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 28-426 of gC from HSV-2 (e.g., strain 333 or UL44), as set forth in the following amino acid sequence: SPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTGPPKTSSEPVRCNRHDPLARYGSRVQIRCRF PNSTRTESRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLIIEE LTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYYPGNRAEFVWFEDGRRVFDPAQIHTQT QENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWFLGD DSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHH (SEQ ID NO: 25).

[0171] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 25. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 25.

[0172] In some embodiments, an HSV-2 gC fragment comprises the following amino acid sequence: SPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTGPPKTSSEPVRCNRHDPLARYGSRVQIRCRF PNSTRTEFRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLIIEE LTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYYPGNRAEFVWFEDGRRVFDPAQIHTQT QENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWFLGD DSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHH (SEQ ID NO: 26).

[0173] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 26. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 26.

[0174] In some embodiments, the HSV-2 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-426 of gC from HSV-2 (e.g., strain 333 or UL44), as set forth in the following amino acid sequence: SASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTGPPKTSSEPVRCNRHDPLARYGSRVQIRC RFPNSTRTESRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLII EELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYYPGNRAEFVWFEDGRRVFDPAQIHT QTQENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWFL GDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHH (SEQ ID NO: 27).

[0175] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 27. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 27.Page 35 of 182 12197519v1

[0176] In some embodiments, an HSV-2 gC fragment comprises the following amino acid sequence: SASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTGPPKTSSEPVRCNRHDPLARYGSRVQIRC RFPNSTRTEFRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLII EELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYYPGNRAEFVWFEDGRRVFDPAQIHT QTQENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWFL GDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHH (SEQ ID NO: 28).

[0177] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 28. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 28.

[0178] In some embodiments, the full-length HSV-2 gC encoded by an RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MALGRVGLAVGLWGLLWVGVVVVLANASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTGPP KTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTEFRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIW AEGAGPGASPRLYSVVGPLGRQRLIIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYY PGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITM EFTGDHAVCTAGCVPEGVTFAWFLGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHHGSHQPPP RDPTERQVIRAVEGAGIGVAVLVAVVLAGTAVVYLTHASSVRYRRLR (SEQ ID NO: 29).

[0179] In some embodiments, an HSV-2 gC comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 29. In some embodiments, an HSV-2 gC has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 29.

[0180] In another embodiment, the HSV-2 gC, or an immunogenic fragment thereof, encoded by an RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA20532.1, AAA66442.1, AAB60549.1, AAB60550.1, AAB60551.1, AAB72101.1, ABU45429.1, ABU45430.1, ABU45431.1, ABU45432.1, ABU45459.1, ABU45460.1, AEV91348.1, AEV91383.1, AEV91407.1, AFM93864.1, AHG54708.1, AKC42808.1, AKC59285.1, AKC59357.1, AKC59428.1, AKC59499.1, AKC59570.1, AMB66008.1, AMB66079.1, AMB66151.1, AMB66224.1, AMB66252.1, AMB66253.1, AMB66368.1, AMB66441.1, AQZ55735.2, AQZ55806.1, AQZ55877.1, AQZ55948.1, AQZ56019.1, AQZ56090.1, AQZ56161.2, AQZ56232.2, AQZ56303.2, AQZ56374.2, AQZ56445.1, AQZ56516.1, AQZ56587.1, AQZ56658.1, AQZ56729.2, AQZ56800.1, AQZ56871.1, AQZ56942.2, AQZ57013.1, AQZ57084.2, AQZ57155.1, AQZ57226.1, AQZ57297.1, AQZ57368.1, AQZ57439.1, AQZ57510.1, AQZ57581.1, AQZ57652.1, AQZ57723.1, AQZ57794.2, AQZ57865.2, AQZ57936.1, AQZ58007.2, AQZ58078.1, AQZ58149.2, AQZ58220.1, AQZ58291.1, AQZ58362.1, AQZ58433.1, AQZ58504.1, AQZ58575.1, AQZ58646.1, AQZ58717.2, AQZ58788.2, AQZ58859.2, AQZ58930.1, AQZ59001.2, AQZ59072.1, AQZ59143.1, ARO38067.1, ARO38068.1, ARO38069.1, ARO38070.1, ARO38071.1, ARO38072.1, CAA25687.1, CAA26025.1, CAB06730.1, CAB06734.1, CAB96544.1, P03173.1, P06475.1, P89475.1, Q89730.1, YP_009137161.1, YP_009137196.1, or YP_009137220.1.Page 36 of 182 12197519v1

[0181] In some embodiments, the gC protein fragment encoded by an RNA utilized in the methods and compositions of the present disclosure comprises a properdin interfering domain. HSV Glycoprotein D

[0182] In some embodiments, compositions of the present disclosure further comprise an RNA or a nucleoside- modified RNA encoding a polypeptide comprising, or consisting of, HSV glycoprotein D (gD), or an immunogenic fragment thereof.

[0183] In some embodiments, disclosed herein is a single composition comprising an RNA encoding an HSV gB and an RNA encoding an HSV gD, or immunogenic fragments thereof. In some embodiments, said gB and said gD RNAs are part of a single polyribonucleotide. In some embodiments, disclosed herein are two individual compositions to be administered as part of a single treatment, the first composition comprising an RNA encoding an HSV gB or an immunogenic fragment thereof, and the second composition comprising an RNA encoding an HSV gD or immunogenic fragment thereof.

[0184] In some embodiments, an RNA encoding an HSV gD, or an immunogenic fragment thereof, comprises an RNA encoding HSV-1 gD, also termed gD1, or an immunogenic fragment thereof. In some embodiments, a nucleotide sequence of the RNA encoding an HSV-1 gD fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAA UCAUUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCUGCUGCUG CUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCAAGUACGCCCUGGCCGACGCCUCCCUGAAGAUGGCCGACCCC AACCGCUUCCGCGGCAAGGACCUGCCCGUGCUGGACCAGCUGACCGACCCCCCCGGCGUGCGCCGCGUGUACCACAUCCAGGC CGGCCUGCCCGACCCCUUCCAGCCCCCCUCCCUGCCCAUCACCGUGUACUACGCCGUGCUGGAGCGCGCCUGCCGCUCCGUGC UGCUGAACGCCCCCUCCGAGGCCCCCCAGAUCGUGCGCGGCGCCUCCGAGGACGUGCGCAAGCAGCCCUACAACCUGACCAUC GCCUGGUUCCGCAUGGGCGGCAACUGCGCCAUCCCCAUCACCGUGAUGGAGUACACCGAGUGCUCCUACAACAAGUCCCUGG GCGCCUGCCCCAUCCGCACCCAGCCCCGCUGGAACUACUACGACUCCUUCUCCGCCGUGUCCGAGGACAACCUGGGCUUCCUG AUGCACGCCCCCGCCUUCGAGACCGCCGGCACCUACCUGCGCCUGGUGAAGAUCAACGACUGGACCGAGAUCACCCAGUUCAU CCUGGAGCACCGCGCCAAGGGCUCCUGCAAGUACGCCCUGCCCCUGCGCAUCCCCCCCUCCGCCUGCCUGUCCCCCCAGGCCU ACCAGCAGGGCGUGACCGUGGACUCCAUCGGCAUGCUGCCCCGCUUCAUCCCCGAGAACCAGCGCACCGUGGCCGUGUACUC CCUGAAGAUCGCCGGCUGGCACGGCCCCAAGGCCCCCUACACCUCCACCCUGCUGCCCCCCGAGCUGUCCGAGACCCCCAACG CCACCCAGCCCGAGCUGGCCCCCGAGGACCCCGAGGACUCCGCCCUGCUGGAGGACCCCGUGGGCACCGUGGCCCCCCAGAUC CCCCCCAACUGGCACAUCCCCUCCAUCCAGGACGCCGCCACCCCCUACUAACUAGUAGUGACUGACUAGGAUCUGGUUACCAC UAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGUUGUCCCCCAAAAU GUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 47)

[0185] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 13). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 38). In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 3) and poly adenylation tail (SEQ ID NO: 15).Page 37 of 182 12197519v1

[0186] In another embodiment, the nucleotide sequence of the RNA encoding an HSV-1 gD fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof. In some embodiments, the polynucleotide sequence of the HSV-1 gD fragment is as set forth in SEQ ID NO: 32.

[0187] In some embodiments, the HSV-1 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-331 of gD (e.g., from HSV-1 Patton strain), as set forth in the following amino acid sequence: KYALADASLKMADPNRFRGKDLPVLDQLTDPPGVRRVYHIQAGLPDPFQPPSLPITVYYAVLERACRSVLLNAPSEAPQIVRGASEDV RKQPYNLTIAWFRMGGNCAIPITVMEYTECSYNKSLGACPIRTQPRWNYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTE ITQFILEHRAKGSCKYALPLRIPPSACLSPQAYQQGVTVDSIGMLPRFIPENQRTVAVYSLKIAGWHGPKAPYTSTLLPPELSETPNAT QPELAPEDPEDSALLEDPVGTVAPQIPPNWHIPSIQDAATPY (SEQ ID NO: 32)

[0188] In some embodiments, an HSV-1 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 32. In some embodiments, an HSV-1 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 32.

[0189] In some embodiments, the full-length HSV-1 gD encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MGGAAARLGAVILFVVIVGLHGVRGKYALADASLKLADPNRFRRKDLPVLDQLTDPPGVRRVYHIQAGLPDPFQPPSLPITVYYAVLE RACRSVLLNAPSEAPQIVRGASEDVRKQPYNLTIAWFRMGGNCAIPITVMEYTECSYNKSLGACPIRTQPRWNYYDSFSAVSEDNLG FLMHAPAFETAGTYLRLVKINDWTEITQFILEHRAKGSCKYALPLRIPPSACLSPQAYQQGVTVDSIGMLPRFIPENQRTVAVYSLKIA GWHGPKAPYTSTLLPPELSETPNATQPELAPEAPEDSALLEDPVGTVAPQIPPNWHIPSIQDAATPYHPPATPNNMGLIAGAVGGSLL AALVICGIVYWMRRRTQKAPKRIRLPHIREDDQPSSHQPLFY (SEQ ID NO: 33)

[0190] In some embodiments, an HSV-1 gD comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 33. In some embodiments, an HSV-1 gD has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 33.

[0191] In another embodiment, the HSV-1 gD, or an immunogenic fragment thereof, encoded by an RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any one of the following GenBank Accession Numbers: AAL90884.1 (KHS2 strain), AAL90883.1 (KHS1 strain), AAK93950.1 (F strain), AAB59754.1 (F strain), AAA19631.1 (mutant strain not identified), AAA19630.1 (mutant strain not identified), or AAA19629.1 (strain not identified).

[0192] In another embodiment, the HSV-1 gD, or an immunogenic fragment thereof, encoded by an RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: A1Z0Q5.2, AAA45780.1, AAA45785.1, AAA45786.1, AAA96682.1, AAK19597.1, AAN74642.1, ABI63524.1, ABM52978.1, ABM52979.1, ABM52980.1, ABM52981.1, ABM66847.1, ABM66848.1, ACM62295.1, ADD60053.1, ADD60130.1, ADM22389.1, ADM22466.1, ADM22542.1, ADM22619.1, ADM22696.1, ADM22773.1, ADM22849.1, ADM22926.1, ADM23003.1, ADM23079.1, ADM23155.1, ADM23231.1,Page 38 of 182 12197519v1ADM23309.1, ADM23383.1, ADM23457.1, ADM23531.1, ADM23605.1, ADM23680.1, ADM23755.1, ADM23831.1, AEQ77097.1, AER37647.1, AER37715.1, AER37786.1, AER37857.1, AER37929.1, AER38000.1, AER38070.1, AFE62894.1, AFH41180.1, AFI23657.1, AFK50415.1, AFP86430.1, AGZ01928.1, AIR95858.1, AJE60009.1, AJE60080.1, AJE60151.1, AJE60222.1, AJE60293.1, AJE60439.1, AKE48645.1, AKG59246.1, AKG59318.1, AKG59391.1, AKG59462.1, AKG59536.1, AKG59609.1, AKG59682.1, AKG59755.1, AKG59826.1, AKG59898.1, AKG59972.1, AKG60046.1, AKG60118.1, AKG60189.1, AKG60261.1, AKG60334.1, AKG60404.1, AKG60474.1, AKG60546.1, AKG60620.1, AKG60692.1, AKG60763.1, AKG60835.1, AKG60906.1, AKG60978.1, AKG61050.1, AKG61123.1, AKG61194.1, AKG61267.1, AKG61339.1, AKG61411.1, AKG61484.1, AKG61556.1, AKG61629.1, AKG61703.1, AKG61774.1, AKG61847.1, AKG61920.1, AKG61993.1, AKH80463.1, AKH80536.1, ALM22635.1, ALM22709.1, ALM22783.1, ALM22857.1, ALO18662.1, ALO18738.1, AMB65662.1, AMB65735.1, AMB65809.1, AMB65885.1, AMB65956.1, AMN09832.1, ANN83964.1, ANN84041.1, ANN84117.1, ANN84194.1, ANN84271.1, ANN84348.1, ANN84424.1, ANN84500.1, ANN84577.1, ANN84653.1, ANN84730.1, ANN84806.1, ANN84883.1, ANN84959.1, ANN85036.1, ANN85112.1, ANN85187.1, ANN85264.1, ANN85341.1, ANN85416.1, ANN85494.1, ANN85571.1, ANN85648.1, ANN85724.1, ANN85801.1, AOY34093.1, AOY34141.1, AOY34243.1, AOY34271.1, AOY34337.1, AOY36685.1, ARB08957.1, ARO37961.1, ARO37962.1, ARO37963.1, ARO37964.1, ARO37965.1, ARO37966.1, ARO37967.1, ARO37968.1, ARO37969.1, ARO37970.1, ARO37971.1, ARO37972.1, ARO37973.1, ARO37974.1, ARO37975.1, ARO37976.1, ARO37977.1, ARO37978.1, ARO37979.1, ARO37980.1, ARO37981.1, ARO37982.1, ARO37983.1, ARO37984.1, ARO37985.1, ARO37986.1, ARO37987.1, ARO37988.1, ARO37989.1, ARO37990.1, ARO37991.1, ARO37992.1, ARO37993.1, ARO37994.1, ARO37995.1, ARO37996.1, ARO37997.1, ARO37998.1, ARO37999.1, ASM47664.1, ASM47741.1, ASM47818.1, ASM47893.1, BAM73419.1, CAA26060.1, CAA32283.1, CAA32284.1, CAA32289.1, CAA38245.1, CAT05431.1, P06476.1, P36318.1, P57083.1, P68331.1, Q05059.1, Q69091.1, SBO07792.1, SBO07819.1, SBO07855.1, SBO07869.1, SBO07887.1, SBO07908.1, SBS69553.1, SBS69561.1, SBS69579.1, SBS69625.1, SBS69688.1, SBS69694.1, SBS69717.1, SBS69727.1, SBS69811.1, SBT69395.1, SCL76902.1, VGBEDZ, or YP_009137141.1.

[0193] In some embodiments, an RNA encoding HSV gD as described herein comprises an RNA encoding HSV-2 gD, also termed gD2, or an immunogenic fragment thereof.

[0194] In some embodiments, the nucleotide sequence of an RNA encoding an HSV-2 gD fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAA UCAUUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGACCCGCCUGACCGUGCUG GCCCUGCUGGCCGGCCUGCUGGCCUCCUCCCGCGCCAAGUACGCCCUGGCCGACCCCUCCCUGAAGAUGGCCGACCCC AACCGCUUCCGCGGCAAGAACCUGCCCGUGCUGGACCAGCUGACCGACCCCCCCGGCGUGAAGCGCGUGUACCACAUCCAGCC CUCCCUGGAGGACCCCUUCCAGCCCCCCUCCAUCCCCAUCACCGUGUACUACGCCGUGCUGGAGCGCGCCUGCCGCUCCGUGC UGCUGCACGCCCCCUCCGAGGCCCCCCAGAUCGUGCGCGGCGCCUCCGACGAGGCCCGCAAGCACACCUACAACCUGACCAUC GCCUGGUACCGCAUGGGCGACAACUGCGCCAUCCCCAUCACCGUGAUGGAGUACACCGAGUGCCCCUACAACAAGUCCCUGG GCGUGUGCCCCAUCCGCACCCAGCCCCGCUGGUCCUACUACGACUCCUUCUCCGCCGUGUCCGAGGACAACCUGGGCUUCCU GAUGCACGCCCCCGCCUUCGAGACCGCCGGCACCUACCUGCGCCUGGUGAAGAUCAACGACUGGACCGAGAUCACCCAGUUCA UCCUGGAGCACCGCGCCCGCGCCUCCUGCAAGUACGCCCUGCCCCUGCGCAUCCCCCCCGCCGCCUGCCUGACCUCCAAGGCC UACCAGCAGGGCGUGACCGUGGACUCCAUCGGCAUGCUGCCCCGCUUCAUCCCCGAGAACCAGCGCACCGUGGCCCUGUACUPage 39 of 182 12197519v1CCCUGAAGAUCGCCGGCUGGCACGGCCCCAAGCCCCCCUACACCUCCACCCUGCUGCCCCCCGAGCUGUCCGACACCACCAAC GCCACCCAGCCCGAGCUGGUGCCCGAGGACCCCGAGGACUCCGCCCUGCUGGAGGACCCCGCCGGCACCGUGUCCUCCCAGA UCCCCCCCAACUGGCACAUCCCCUCCAUCCAGGACGUGGCCCCCCACCACUAACUAGUAGUGACUGACUAGGAUCUGGUUACC ACUAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGUUGUCCCCCAAAA UGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 34)

[0195] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 13). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 38). In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 15) and poly adenylation tail (SEQ ID NO: 16).

[0196] In some embodiments, a polyribonucleotide encoding a gD2 signal sequence comprises the sequence ATGACCCGCCTGACCGTGCTGGCCCTGCTGGCCGGCCTGCTGGCCTCCTCCCGCGCC (SEQ ID NO: 38)

[0197] In another embodiment, the nucleotide sequence of the RNA encoding an HSV-2 gD fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof. In some embodiments, the nucleotide sequence of the RNA encoding an HSV-2 gD fragment is as set forth in SEQ ID NO: 35.

[0198] In some embodiments, the HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-331 of gD (e.g., from HSV-2 strain 333 or US6), as set forth in the following amino acid sequence: KYALADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLERACRSVLLHAPSEAPQIVRGASDEAR KHTYNLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEI TQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQ PELVPEDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHH (SEQ ID NO: 36).

[0199] In some embodiments, an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 36. In some embodiments, an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 36.

[0200] In some embodiments, an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 30-331 of gD from HSV-2(e.g., strain 333 or US6), as set forth in the following amino acid sequence: ADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLERACRSVLLHAPSEAPQIVRGASDEARKHT YNLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEITQF ILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPEL VPEDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHH (SEQ ID NO: 39).

[0201] In some embodiments, an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at leastPage 40 of 182 12197519v198%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 39. In some embodiments, an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 39.

[0202] In some embodiments, an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 31-331 of gD from HSV-2 (e.g., strain 333 or US6), as set forth in the following amino acid sequence: DPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLERACRSVLLHAPSEAPQIVRGASDEARKHTY NLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEITQFIL EHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVP EDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHH (SEQ ID NO: 40).

[0203] In some embodiments, an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 40. In some embodiments, an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 40.

[0204] In some embodiments, the full-length HSV-2 gD encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MGRLTSGVGTAALLVVAVGLRVVCAKYALADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLE RACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLG FLMHAPAFETAGTYLRLVKINDWTEITQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYSLKIA GWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHHAPAAPSNPGLIIGALAGSTLA VLVIGGIAFWVRRRAQMAPKRLRLPHIRDDDAPPSHQPLFY (SEQ ID NO: 37).

[0205] In some embodiments, an HSV-2 gD comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 37. In some embodiments, an HSV-2 gD has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 37.

[0206] In another embodiment, the HSV-2 gD, or immunogenic fragment thereof, encoded by AN RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in GenBank Accession Numbers: 1003204A, AAA45841.1, AAA45842.1, AAB60552.1, AAB60553.1, AAB60554.1, AAB60555.1, AAB72102.1, AAS01730.1, AAW23130.1, AAW23131.1, AAW23132.1, AAW23133.1, AAW23134.1, ABS84899.1, ABU45433.1, ABU45434.1, ABU45435.1, ABU45461.1, ABU45462.1, ACA28831.1, AEV91405.1, AFM93876.1, AFS18198.1, AFS18199.1, AFS18200.1, AFS18201.1, AFS18202.1, AFS18203.1, AFS18204.1, AFS18205.1, AFS18206.1, AFS18207.1, AFS18208.1, AFS18209.1, AFS18210.1, AFS18211.1, AFS18212.1, AFS18213.1, AFS18214.1, AFS18215.1, AFS18216.1, AFS18217.1, AFS18218.1, AFS18219.1, AFS18220.1, AFS18221.1, AHG54730.1, AIL27720.1, AIL27721.1, AIL27722.1, AIL27723.1, AIL27724.1, AIL27725.1, AIL27726.1, AIL27727.1, AIL27728.1, AIL27729.1, AIL27730.1, AIL27731.1, AIL28069.1, AIL28070.1, AKC42828.1, AKC59305.1, AKC59376.1, AKC59447.1, AKC59518.1, AKC59589.1, AMB66102.1, AMB66171.1, AMB66244.1, AMB66321.1, AMB66394.1, AMB66463.1, AQZ55754.1, AQZ55825.1, AQZ55896.1, AQZ55967.1, AQZ56038.1, AQZ56109.1, AQZ56180.1, AQZ56251.1, AQZ56322.1, AQZ56393.1, AQZ56464.1, AQZ56535.1, AQZ56606.1, AQZ56677.1, AQZ56748.1,Page 41 of 182 12197519v1AQZ56819.1, AQZ56890.1, AQZ56961.1, AQZ57032.1, AQZ57103.1, AQZ57174.1, AQZ57245.1, AQZ57316.1, AQZ57387.1, AQZ57458.1, AQZ57529.1, AQZ57600.1, AQZ57671.1, AQZ57742.1, AQZ57813.1, AQZ57884.1, AQZ57955.1, AQZ58026.1, AQZ58097.1, AQZ58168.1, AQZ58239.1, AQZ58310.1, AQZ58381.1, AQZ58452.1, AQZ58523.1, AQZ58594.1, AQZ58665.1, AQZ58736.1, AQZ58807.1, AQZ58878.1, AQZ58949.1, AQZ59020.1, AQZ59091.1, AQZ59162.1, ARO38000.1, ARO38001.1, ARO38002.1, ARO38003.1, ARO38004.1, ARO38005.1, ARO38006.1, ARO38007.1, ARO38008.1, ARO38009.1, ARO38010.1, ARO38011.1, ARO38012.1, ARO38013.1, ARO38014.1, ARO38015.1, ARO38016.1, ARO38017.1, ARO38018.1, ARO38019.1, ARO38020.1, ARO38021.1, ARO38022.1, ARO38023.1, ARO38024.1, ARO38025.1, ARO38026.1, ARO38027.1, ARO38028.1, ARO38029.1, ARO38030.1, ARO38031.1, ARO38032.1, ARO38033.1, ARO38034.1, ARO38035.1, ARO38036.1, ARO38037.1, ARO38038.1, ARO38039.1, ARO38040.1, ARO38041.1, ARO38042.1, ARO38043.1, ARO38044.1, CAA26025.1, CAB06713.1, CAC33573.1, CAT05432.1, P03172.2, Q69467.1, or YP_009137218.1.

[0207] In some embodiments, the gD protein or fragment (e.g., immunogenic fragment) includes Y63. In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes R159. In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes D240. In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes P246. In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes a residue selected from Y63, R159, D240, and P246. In another embodiment, inclusion of one of these residues elicits antibodies that inhibit binding to nectin-1. HSV Glycoprotein E

[0208] In some embodiments, compositions of the present disclosure further comprise an RNA or a nucleoside- modified RNA encoding a polypeptide comprising, or consisting of, HSV glycoprotein E (gE), or an immunogenic fragment thereof.

[0209] In some embodiments, disclosed herein is a single composition comprising an RNA encoding an HSV gB and an RNA encoding an HSV gE, or immunogenic fragments thereof. In some embodiments, said gB and said gE RNAs are part of a single polyribonucleotide. In some embodiments, disclosed herein are two individual compositions to be administered as part of a single treatment, the first composition comprising an RNA encoding an HSV gB or an immunogenic fragment thereof, and the second composition comprising an RNA encoding an HSV gE or immunogenic fragment thereof.

[0210] In some embodiments, an RNA encoding an HSV gE, or an immunogenic fragment thereof, comprises an RNA encoding HSV-1 gE, also termed gE1, or an immunogenic fragment thereof. In some embodiments, a nucleotide sequence of the RNA encoding an HSV-1 gE fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAA UCAUUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCUGCUGCUG CUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCAAGACCUCCUGGCGCCGCGUGUCCGUGGGCGAGGACGUGUCC CUGCUGCCCGCCCCCGGCCCCACCGGCCGCGGCCCCACCCAGAAGCUGCUGUGGGCCGUGGAGCCCCUGGACGGCUGCGGCC CCCUGCACCCCUCCUGGGUGUCCCUGAUGCCCCCCAAGCAGGUGCCCGAGACCGUGGUGGACGCCGCCUGCAUGCGCGCCCC CGUGCCCCUGGCCAUGGCCUACGCCCCCCCCGCCCCCUCCGCCACCGGCGGCCUGCGCACCGACUUCGUGUGGCAGGAGCGC GCCGCCGUGGUGAACCGCUCCCUGGUGAUCUACGGCGUGCGCGAGACCGACUCCGGCCUGUACACCCUGUCCGUGGGCGACA UCAAGGACCCCGCCCGCCAGGUGGCCUCCGUGGUGCUGGUGGUGCAGCCCGCCCCCGUGCCCACCCCCCCCCCCACCCCCGCCPage 42 of 182 12197519v1GACUACGACGAGGACGACAACGACGAGGGCGAGGGCGAGGACGAGUCCCUGGCCGGCACCCCCGCCUCCGGCACCCCCCGCCU GCCCCCCUCCCCCGCCCCCCCCCGCUCCUGGCCCUCCGCCCCCGAGGUGUCCCACGUGCGCGGCGUGACCGUGCGCAUGGAGA CCCCCGAGGCCAUCCUGUUCUCCCCCGGCGAGGCCUUCUCCACCAACGUGUCCAUCCACGCCAUCGCCCACGACGACCAGACC UACACCAUGGACGUGGUGUGGCUGCGCUUCGACGUGCCCACCUCCUGCGCCGAGAUGCGCAUCUACGAGUCCUGCCUGUACC ACCCCCAGCUGCCCGAGUGCCUGUCCCCCGCCGACGCCCCCUGCGCCGCCUCCACCUGGACCUCCCGCCUGGCCGUGCGCUCC UACGCCGGCUGCUCCCGCACCAACCCCCCCCCCCGCUGCUCCGCCGAGGCCCACAUGGAGCCCUUCCCCGGCCUGGCCUGGCA GGCCGCCUCCGUGAACCUGGAGUUCCGCGACGCCUCCCCCCAGCACUCCGGCCUGUACCUGUGCGUGGUGUACGUGAACGAC CACAUCCACGCCUGGGGCCACAUCACCAUCAACACCGCCGCCCAGUACCGCAACGCCGUGGUGGAGCAGCCCCUGCCCCAGCG CGGCGCCGACCUGGCCGAGCCCACCCACCCCCACGUGGGCGCCUAACUAGUAGUGACUGACUAGGAUCUGGUUACCACUAAAC CAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGUUGUCCCCCAAAAUGUAGC CAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 41).

[0211] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 13). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 3). In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 15) and poly adenylation tail (SEQ ID NO: 16).

[0212] In another embodiment, the nucleotide sequence of the RNA encoding an HSV-1 gE fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof. In some embodiments, the sequence of the HSV-1 gE fragment is as set forth in SEQ ID NO: 42.

[0213] In some embodiments, the HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-409 of gE from HSV-1 (e.g., NS strain), as set forth in the following amino acid sequence: KTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPPKQVPETVVDAACMRAPVPLAMAYAPPAPSATG GLRTDFVWQERAAVVNRSLVIYGVRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPTPADYDEDDNDEGEGEDESLAGT PASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNVSIHAIAHDDQTYTMDVVWLRFDVPTSCAEMRIYES CLYHPQLPECLSPADAPCAASTWTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLAWQAASVNLEFRDASPQHSGLYLCVVYVND HIHAWGHITINTAAQYRNAVVEQPLPQRGADLAEPTHPHVGA (SEQ ID NO: 43).

[0214] In some embodiments, an HSV-1 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 43. In some embodiments, an HSV-1 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 43. In some embodiments, the gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-409 of gE from an HSV-1 strain (e.g., SEQ ID NO: 43).

[0215] In some embodiments, the HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 21-409 of gE from HSV-1 (e.g., NS strain or US8), as set forth in the following amino acid sequence:Page 43 of 182 12197519v1GTPKTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPPKQVPETVVDAACMRAPVPLAMAYAPPAPSA TGGLRTDFVWQERAAVVNRSLVIYGVRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPTPADYDEDDNDEGEGEDESLA GTPASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNVSIHAIAHDDQTYTMDVVWLRFDVPTSCAEMRIY ESCLYHPQLPECLSPADAPCAASTWTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLAWQAASVNLEFRDASPQHSGLYLCVVYVN DHIHAWGHITINTAAQYRNAVVEQPLPQRGADLAEPTHPHVGA (SEQ ID NO: 44).

[0216] In some embodiments, an HSV-1 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 44. In some embodiments, an HSV-1 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 44. In some embodiments, the gE fragment encoded by an RNA utilized in the methods and compositions of the present disclosure comprises amino acids 21-409 of gE from an HSV-1 strain (e.g., SEQ ID NO: 44).

[0217] In some embodiments, the full-length HSV-1 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MDRGAVVGFLLGVCVVSCLAGTPKTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPPKQVPETVVD AACMRAPVPLAMAYAPPAPSATGGLRTDFVWQERAAVVNRSLVIYGVRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPT PADYDEDDNDEGEGEDESLAGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNVSIHAIAHDDQTYT MDVVWLRFDVPTSCAEMRIYESCLYHPQLPECLSPADAPCAASTWTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLAWQAASVN LEFRDASPQHSGLYLCVVYVNDHIHAWGHITINTAAQYRNAVVEQPLPQRGADLAEPTHPHVGAPPHAPPTHGALRLGAVMGAALL LSALGLSVWACMTCWRRRAWRAVKSRASGKGPTYIRVADSELYADWSSDSEGERDQVPWLAPPERPDSPSTNGSGFEILSPTAPSV YPRSDGHQSRRQLTTFGSGRPDRRYSQASDSSVFW (SEQ ID NO: 45).

[0218] In some embodiments, an HSV-1 gE comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 45. In some embodiments, an HSV-1 gE has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 45.

[0219] In another embodiment, the HSV-1 gE, or an immunogenic fragment thereof, encoded by an RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA45779.1, AAA96680.1, ABI63526.1, ACM62297.1, ADD60055.1, ADD60132.1, ADM22391.1, ADM22468.1, ADM22544.1, ADM22621.1, ADM22698.1, ADM22775.1, ADM22851.1, ADM22928.1, ADM23005.1, ADM23081.1, ADM23157.1, ADM23233.1, ADM23311.1, ADM23385.1, ADM23459.1, ADM23533.1, ADM23607.1, ADM23682.1, ADM23757.1, ADM23833.1, ADN34689.1, ADN34692.1, ADN34695.1, AEQ77099.1, AER37649.1, AER37717.1, AER37788.1, AER37859.1, AER37931.1, AER38002.1, AER38072.1, AFA36179.1, AFA36180.1, AFA36181.1, AFA36182.1, AFA36183.1, AFA36184.1, AFA36185.1, AFA36186.1, AFA36187.1, AFA36188.1, AFA36189.1, AFA36190.1, AFA36191.1, AFA36192.1, AFA36193.1, AFA36194.1, AFA36195.1, AFA36196.1, AFA36197.1, AFA36198.1, AFA36199.1, AFA36200.1, AFA36201.1, AFA36202.1, AFA36203.1, AFE62896.1, AFI23659.1, AFK50417.1, AFP86432.1, AGZ01930.1, AIR95859.1, AJE60011.1, AJE60082.1, AJE60153.1, AJE60224.1, AJE60295.1, AKE48647.1, AKE98373.1, AKE98374.1, AKE98375.1, AKE98376.1, AKE98377.1, AKE98378.1, AKE98379.1, AKE98380.1, AKE98381.1, AKE98382.1, AKE98383.1, AKE98384.1, AKE98385.1, AKE98386.1, AKE98387.1, AKE98388.1, AKE98389.1, AKE98390.1, AKE98391.1,Page 44 of 182 12197519v1AKE98392.1, AKE98393.1, AKG59248.1, AKG59320.1, AKG59393.1, AKG59464.1, AKG59538.1, AKG59611.1, AKG59684.1, AKG59757.1, AKG59828.1, AKG59900.1, AKG59974.1, AKG60048.1, AKG60120.1, AKG60191.1, AKG60263.1, AKG60336.1, AKG60406.1, AKG60476.1, AKG60548.1, AKG60622.1, AKG60694.1, AKG60765.1, AKG60837.1, AKG60908.1, AKG60980.1, AKG61052.1, AKG61125.1, AKG61196.1, AKG61269.1, AKG61341.1, AKG61413.1, AKG61486.1, AKG61558.1, AKG61631.1, AKG61705.1, AKG61776.1, AKG61849.1, AKG61922.1, AKG61995.1, AKH80465.1, AKH80538.1, ALM22637.1, ALM22711.1, ALM22785.1, ALM22859.1, ALO18664.1, ALO18740.1, AMB65664.1, AMB65737.1, AMB65811.1, AMB65887.1, AMB65958.1, AMN09834.1, ANN83966.1, ANN84043.1, ANN84119.1, ANN84196.1, ANN84273.1, ANN84350.1, ANN84426.1, ANN84502.1, ANN84579.1, ANN84655.1, ANN84732.1, ANN84808.1, ANN84885.1, ANN84961.1, ANN85038.1, ANN85114.1, ANN85189.1, ANN85266.1, ANN85343.1, ANN85418.1, ANN85496.1, ANN85573.1, ANN85650.1, ANN85726.1, ANN85803.1, AOY34085.1, AOY36687.1, ARB08959.1, ARO38073.1, ARO38074.1, ARO38075.1, ARO38076.1, ARO38077.1, ARO38078.1, ARO38079.1, ARO38080.1, ASM47642.1, ASM47666.1, ASM47743.1, ASM47820.1, ASM47895.1, BAM73421.1, CAA26062.1, CAA32272.1, CAF24756.1, CAF24757.1, CAF24758.1, CAF24759.1, CAF24760.1, CAF24761.1, CAF24762.1, CAF24763.1, CAF24764.1, CAF24765.1, CAF24766.1, CAF24767.1, CAF24768.1, CAF24769.1, CAF24770.1, CAF24771.1, CAF24772.1, CAF24773.1, CAF24774.1, CAF24775.1, CAF24776.1, CAF24777.1, CAF24778.1, CAF24779.1, CAF24780.1, CAF24781.1, CAF24782.1, CAF24783.1, CAF24784.1, CAF24785.1, P04290.1, P04488.1, P28986.1, Q703F0.1, SBO07910.1, SBS69571.1, SBS69576.1, SBS69595.1, SBS69636.1, SBS69693.1, SBS69701.1, SBS69722.1, SBS69732.1, SBS69813.1, SBT69397.1, or YP_009137143.1.

[0220] In some embodiments, an RNA encoding HSV gE as described herein comprises an RNA encoding HSV-2 gE, also termed gE2, or an immunogenic fragment thereof.

[0221] In some embodiments, a nucleotide sequence of an RNA encoding an HSV-2 gE fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAA UCAUUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCUGCUGCUG CUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCCGCACCUCCUGGAAGCGCGUGACCUCCGGCGAGGACGUGGUG CUGCUGCCCGCCCCCGCCGGCCCCGAGGAGCGCACCCGCGCCCACAAGCUGCUGUGGGCCGCCGAGCCCCUGGACGCCUGCG GCCCCCUGCGCCCCUCCUGGGUGGCCCUGUGGCCCCCCCGCCGCGUGCUGGAGACCGUGGUGGACGCCGCCUGCAUGCGCGC CCCCGAGCCCCUGGCCAUCGCCUACUCCCCCCCCUUCCCCGCCGGCGACGAGGGCCUGUACUCCGAGCUGGCCUGGCGCGACC GCGUGGCCGUGGUGAACGAGUCCCUGGUGAUCUACGGCGCCCUGGAGACCGACUCCGGCCUGUACACCCUGUCCGUGGUGG GCCUGUCCGACGAGGCCCGCCAGGUGGCCUCCGUGGUGCUGGUGGUGGAGCCCGCCCCCGUGCCCACCCCCACCCCCGACGA CUACGACGAGGAGGACGACGCCGGCGUGUCCGAGCGCACCCCCGUGUCCGUGCCCCCCCCCACCCCCCCCCGCCGCCCCCCCG UGGCCCCCCCCACCCACCCCCGCGUGAUCCCCGAGGUGUCCCACGUGCGCGGCGUGACCGUGCACAUGGAGACCCCCGAGGCC AUCCUGUUCGCCCCCGGCGAGACCUUCGGCACCAACGUGUCCAUCCACGCCAUCGCCCACGACGACGGCCCCUACGCCAUGGA CGUGGUGUGGAUGCGCUUCGACGUGCCCUCCUCCUGCGCCGAGAUGCGCAUCUACGAGGCCUGCCUGUACCACCCCCAGCUG CCCGAGUGCCUGUCCCCCGCCGACGCCCCCUGCGCCGUGUCCUCCUGGGCCUACCGCCUGGCCGUGCGCUCCUACGCCGGCU GCUCCCGCACCACCCCCCCCCCCCGCUGCUUCGCCGAGGCCCGCAUGGAGCCCGUGCCCGGCCUGGCCUGGCUGGCCUCCACC GUGAACCUGGAGUUCCAGCACGCCUCCCCCCAGCACGCCGGCCUGUACCUGUGCGUGGUGUACGUGGACGACCACAUCCACG CCUGGGGCCACAUGACCAUCUCCACCGCCGCCCAGUACCGCAACGCCGUGGUGGAGCAGCACCUGCCCCAGCGCCAGCCCGAG CCCGUGGAGCCCACCCGCCCCCACGUGCGCGCCUAACUAGUAGUGACUGACUAGGAUCUGGUUACCACUAAACCAGCCUCAAGPage 45 of 182 12197519v1AACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUC UGCUCCUAAUAAAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 46).

[0222] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 13). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 3). In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 15) and poly adenylation tail (SEQ ID NO: 16).

[0223] In another embodiment, the nucleotide sequence of the RNA encoding an HSV-2 gE fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof. In some embodiments, the sequence of the HSV-2 gE fragment is as set forth in SEQ ID NO: 48.

[0224] In some embodiments, the HSV-2 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-405 of gE from HSV-2 (e.g., strain 2.12 or US8) as set forth in the following amino acid sequence: RTSWKRVTSGEDVVLLPAPAGPEERTRAHKLLWAAEPLDACGPLRPSWVALWPPRRVLETVVDAACMRAPEPLAIAYSPPFPAGDE GLYSELAWRDRVAVVNESLVIYGALETDSGLYTLSVVGLSDEARQVASVVLVVEPAPVPTPTPDDYDEEDDAGVSERTPVSVPPPTPP RRPPVAPPTHPRVIPEVSHVRGVTVHMETPEAILFAPGETFGTNVSIHAIAHDDGPYAMDVVWMRFDVPSSCAEMRIYEACLYHPQL PECLSPADAPCAVSSWAYRLAVRSYAGCSRTTPPPRCFAEARMEPVPGLAWLASTVNLEFQHASPQHAGLYLCVVYVDDHIHAWGH MTISTAAQYRNAVVEQHLPQRQPEPVEPTRPHVRA (SEQ ID NO: 48).

[0225] In some embodiments, an HSV-2 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 48. In some embodiments, an HSV-2 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 48.

[0226] In some embodiments, the full-length HSV-2 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MARGAGLVFFVGVWVVSCLAAAPRTSWKRVTSGEDVVLLPAPAERTRAHKLLWAAEPLDACGPLRPSWVALWPPRRVLETVVDAA CMRAPEPLAIAYSPPFPAGDEGLYSELAWRDRVAVVNESLVIYGALETDSGLYTLSVVGLSDEARQVASVVLVVEPAPVPTPTPDDYD EEDDAGVTNARRSAFPPQPPPRRPPVAPPTHPRVIPEVSHVRGVTVHMETLEAILFAPGETFGTNVSIHAIAHDDGPYAMDVVWMRF DVPSSCADMRIYEACLYHPQLPECLSPADAPCAVSSWAYRLAVRSYAGCSRTTPPPRCFAEARMEPVPGLAWLASTVNLEFQHASPQ HAGLYLCVVYVDDHIHAWGHMTISTAAQYRNAVVEQHLPQRQPEPVEPTRPHVRAPHPAPSARGPLRLGAVLGAALLLAALGLSAW ACMTCWRRRSWRAVKSRASATGPTYIRVADSELYADWSSDSEGERDGSLWQDPPERPDSPSTNGSGFEILSPTAPSVYPHSEGRKS RRPLTTFGSGSPGRRHSQASYPSVLW (SEQ ID NO: 49).

[0227] In some embodiments, an HSV-2 gE comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 49. In some embodiments, an HSV-2 gE has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 49.Page 46 of 182 12197519v1

[0228] In another embodiment, the HSV-2 gE, or an immunogenic fragment thereof, encoded by an RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: ABU45436.1, ABU45437.1, ABU45438.1, ABU45439.1, ABW83306.1, ABW83308.1, ABW83310.1, ABW83312.1, ABW83314.1, ABW83316.1, ABW83318.1, ABW83320.1, ABW83322.1, ABW83324.1, ABW83326.1, ABW83328.1, ABW83330.1, ABW83332.1, ABW83334.1, ABW83336.1, ABW83338.1, ABW83340.1, ABW83342.1, ABW83344.1, ABW83346.1, ABW83348.1, ABW83350.1, ABW83352.1, ABW83354.1, ABW83356.1, ABW83358.1, ABW83360.1, ABW83362.1, ABW83364.1, ABW83366.1, ABW83368.1, ABW83370.1, ABW83372.1, ABW83374.1, ABW83376.1, ABW83378.1, ABW83380.1, ABW83382.1, ABW83384.1, ABW83386.1, ABW83388.1, ABW83390.1, ABW83392.1, ABW83394.1, ABW83396.1, ABW83398.1, ABW83400.1, ABZ04069.1, AEV91407.1, AHG54732.1, AKC42830.1, AKC59307.1, AKC59378.1, AKC59449.1, AKC59520.1, AKC59591.1, AMB66104.1, AMB66173.1, AMB66246.1, AMB66465.1, AQZ55756.1, AQZ55827.1, AQZ55898.1, AQZ55969.2, AQZ56040.2, AQZ56111.2, AQZ56182.1, AQZ56253.2, AQZ56324.1, AQZ56395.1, AQZ56466.2, AQZ56537.1, AQZ56608.1, AQZ56679.1, AQZ56750.1, AQZ56821.2, AQZ56892.1, AQZ56963.2, AQZ57034.2, AQZ57105.1, AQZ57176.1, AQZ57247.2, AQZ57318.2, AQZ57389.2, AQZ57460.2, AQZ57531.2, AQZ57602.2, AQZ57673.1, AQZ57744.2, AQZ57815.1, AQZ57886.1, AQZ57957.2, AQZ58028.2, AQZ58099.1, AQZ58170.2, AQZ58241.2, AQZ58312.2, AQZ58383.2, AQZ58454.2, AQZ58525.2, AQZ58596.1, AQZ58667.1, AQZ58738.2, AQZ58809.2, AQZ58880.2, AQZ58951.2, AQZ59022.2, AQZ59093.1, AQZ59164.1, ARO38081.1, ARO38082.1, ARO38083.1, ARO38084.1, ARO38085.1, ARO38086.1, CAB06715.1, P89436.1, P89475.1, or YP_009137220.1. HSV Glycoprotein H

[0229] In some embodiments, compositions of the present disclosure further comprise an RNA or nucleoside- modified RNA encoding HSV glycoprotein H (gH), or an immunogenic fragment thereof. In some embodiments, an RNA encoding an HSV gH, or an immunogenic fragment thereof, comprises an RNA encoding HSV-1 gH, or an immunogenic fragment thereof. In some embodiments, an RNA encoding an HSV gH, or an immunogenic fragment thereof, comprises an RNA encoding HSV-2 gH, or an immunogenic fragment thereof.

[0230] In some embodiments, disclosed herein is a single composition comprising an RNA encoding an HSV gB and an RNA encoding an HSV gH, or immunogenic fragments thereof. In some embodiments, said gB and said gH RNAs are part of a single polyribonucleotide. In some embodiments, disclosed herein are two individual compositions to be administered as part of a single treatment, the first composition comprising an RNA encoding an HSV gB or an immunogenic fragment thereof, and the second composition comprising an RNA encoding an HSV gH or immunogenic fragment thereof.

[0231] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, some uridine residues are 1-methyl-pseudouridine. In another embodiment, the nucleotide sequence of the RNA encoding an HSV-1 or HSV-2 gH fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof.

[0232] In some embodiments, a polyribonucleotide encoding an exemplary polypeptide comprising an HSV-2 gH glycoprotein or an immunogenic fragment thereof, comprises or consists of SEQ ID NO: 189.Page 47 of 182 12197519v1

[0233] In other embodiments, a polyribonucleotide encoding an exemplary polypeptide comprising an HSV-2 gH glycoprotein or immunogenic fragment thereof, comprises or consists of the following nucleotide sequence: CACGACACCTACTGGACCGAGCAGATCGACCCCTGGTTCCTGCACGGCCTGGGCCTGGCCCGCACCTACTGGCGCGACACCAACAC CGGCCGCCTGTGGCTGCCCAACACCCCCGACGCCTCCGACCCCCAGCGCGGCCGCCTGGCCCCCCCCGGCGAGCTGAACCTGACCA CCGCCTCCGTGCCCATGCTGCGCTGGTACGCCGAGCGCTTCTGCTTCGTGCTGGTGACCACCGCCGAGTTCCCCCGCGACCCCGGC CAGCTGCTGTACATCCCCAAGACCTACCTGCTGGGCCGCCCCCGCAACGCCTCCCTGCCCGAGCTGCCCGAGGCCGGCCCCACCTCC CGCCCCCCCGCCGAGGTGACCCAGCTGAAGGGCCTGTCCCACAACCCCGGCGCCTCCGCCCTGCTGCGCTCCCGCGCCTGGGTGAC CTTCGCCGCCGCCCCCGACCGCGAGGGCCTGACCTTCCCCCGCGGCGACGACGGCGCCACCGAGCGCCACCCCGACGGCCGCCGCA ACGCCCCCCCCCCCGGCCCCCCCGCCGGCACCCCCCGCCACCCCACCACCAACCTGTCCATCGCCCACCTGCACAACGCCTCCGTGA CCTGGCTGGCCGCCCGCGGCCTGCTGCGCACCCCCGGCCGCTACGTGTACCTGTCCCCCTCCGCCTCCACCTGGCCCGTGGGCGTG TGGACCACCGGCGGCCTGGCCTTCGGCTGCGACGCCGCCCTGGTGCGCGCCCGCTACGGCAAGGGCTTCATGGGCCTGGTGATCT CCATGCGCGACTCCCCCCCCGCCGAGATCATCGTGGTGCCCGCCGACAAGACCCTGGCCCGCGTGGGCAACCCCACCGACGAGAAC GCCCCCGCCGTGCTGCCCGGCCCCCCCGCCGGCCCCCGCTACCGCGTGTTCGTGCTGGGCGCCCCCACCCCCGCCGACAACGGCTC CGCCCTGGACGCCCTGCGCCGCGTGGCCGGCTACCCCGAGGAGTCCACCAACTACGCCCAGTACATGTCCCGCGCCTACGCCGAGT TCCTGGGCGAGGACCCCGGCTCCGGCACCGACGCCCGCCCCTCCCTGTTCTGGCGCCTGGCCGGCCTGCTGGCCTCCTCCGGCTTC GCCTTCGTGAACGCCGCCCACGCCCACGACGCCATCCGCCTGTCCGACCTGCTGGGCTTCCTGGCCCACTCCCGCGTGCTGGCCGG CCTGGCCGCCCGCGGCGCCGCCGGCTGCGCCGCCGACTCCGTGTTCCTGAACGTGTCCGTGCTGGACCCCGCCGCCCGCCTGCGCC TGGAGGCCCGCCTGGGCCACCTGGTGGCCGCCATCCTGGAGCGCGAGCAGTCCCTGGCCGCCCACGCCCTGGGCTACCAGCTGGC CTTCGTGCTGGACTCCCCCGCCGCCTACGGCGCCGTGGCCCCCTCCGCCGCCCGCCTGATCGACGCCCTGTACGCCGAGTTCCTGG GCGGCCGCGCCCTGACCGCCCCCATGGTGCGCCGCGCCCTGTTCTACGCCACCGCCGTGCTGCGCGCCCCCTTCCTGGCCGGCGCC CCCTCCGCCGAGCAGCGCGAGCGCGCCCGCCGCGGCCTGCTGATCACCACCGCCCTGTGCACCTCCGACGTGGCCGCCGCCACCCA CGCCGACCTGCGCGCCGCCCTGGCCCGCACCGACCACCAGAAGAACCTGTTCTGGCTGCCCGACCACTTCTCCCCCTGCGCCGCCTC CCTGCGCTTCGACCTGGCCGAGGGCGGCTTCATCCTGGACGCCCTGGCCATGGCCACCCGCTCCGACATCCCCGCCGACGTGATGG CCCAGCAGACCCGCGGCGTGGCCTCCGTGCTGACCCGCTGGGCCCACTACAACGCCCTGATCCGCGCCTTCGTGCCCGAGGCCACC CACCAGTGCTCCGGCCCCTCCCACAACGCCGAGCCCCGCATCCTGGTGCCCATCACCCACAACGCCTCCTACGTGGTGACCCACACC CCCCTGCCCCGCGGCATCGGCTACAAGCTGACCGGCGTGGACGTGCGCCGCCCCCTGTTCATCACCTACCTGACCGCCACCTGCGA GGGCCACGCCCGCGAGATCGAGCCCAAGCGCCTGGTGCGCACCGAGAACCGCCGCGACCTGGGCCTGGTGGGCGCCGTGTTCCTG CGCTACACCCCCGCCGGCGAGGTGATGTCCGTGCTGCTGGTGGACACCGACGCCACCCAGCAGCAGCTGGCCCAGGGCCCCGTGGC CGGCACCCCCAACGTGTTCTCCTCCGACGTGCCCTCCGTGGCCCTGCTGCTGTTCCCCAACGGCACCGTGATCCACCTGCTGGCCTT CGACACCCTGCCCATCGCCACCATCGCCCCC (SEQ ID NO: 189).

[0234] In some embodiments, a polyribonucleotide encoding an exemplary polypeptide comprising a gH glycoprotein or an immunogenic fragment thereof, comprises a homology greater than 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 189 or SEQ ID NO: 189. HSV Glycoprotein L

[0235] In some embodiments, compositions of the present disclosure further comprise an RNA or a nucleoside- modified RNA encoding HSV glycoprotein L (gL), or an immunogenic fragment thereof. In some embodiments, an RNA encoding an HSV gL, or an immunogenic fragment thereof, comprises an RNA encoding HSV-1 gL, or anPage 48 of 182 12197519v1immunogenic fragment thereof. In some embodiments, an RNA encoding an HSV gL, or an immunogenic fragment thereof, comprises an RNA encoding HSV-2 gL, or an immunogenic fragment thereof.

[0236] In some embodiments, disclosed herein is a single composition comprising an RNA encoding an HSV gB and an RNA encoding an HSV gL, or immunogenic fragments thereof. In some embodiments, said gB and said gL RNAs are part of a single polyribonucleotide. In some embodiments, disclosed herein are two individual compositions to be administered as part of a single treatment, the first composition comprising an RNA encoding an HSV gB or an immunogenic fragment thereof, and the second composition comprising an RNA encoding an HSV gL or immunogenic fragment thereof.

[0237] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, some uridine residues are 1-methyl-pseudouridine. In another embodiment, the nucleotide sequence of the RNA encoding an HSV-1 or HSV-2 gL fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof.

[0238] In some embodiments, a polyribonucleotide encoding an exemplary polypeptide comprising an HSV-2 gL glycoprotein or an immunogenic fragment thereof, comprises or consists of the following nucleotide sequence: AGCATAAAAGTCTCAACACAACATATACAAAACAAACGAATCTCAAGCAATCAAGCATTCTACTTCTATTGCAGCAATTTAAATCA TTTCTTTTAAAGCAAAAGCAATTTTCTGAAAATTTTCACCATTTACGAACGATAGCGCTATGCGCATGCAGCTGCTGCTGCTGAT CGCCCTGTCCCTGGCCCTGGTGACCAACTCCGGCTCCCAGGCCACCGAGTACGTGCTGCGCTCCGTGATCGCCAAGGAGGTGG GCGACATCCTGCGCGTGCCCTGCATGCGCACCCCCGCCGACGACGTGTCCTGGCGCTACGAGGCCCCCTCCGTGATCGACTACG CCCGCATCGACGGCATCTTCCTGCGCTACCACTGCCCCGGCCTGGACACCTTCCTGTGGGACCGCCACGCCCAGCGCGCCTACC TGGTGAACCCCTTCCTGTTCGCCGCCGGCTTCCTGGAGGACCTGTCCCACTCCGTGTTCCCCGCCGACACCCAGGAGACCACCA CCCGCCGCGCCCTGTACAAGGAGATCCGCGACGCCCTGGGCTCCCGCAAGCAGGCCGTGTCCCACGCCCCCGTGCGCGCCGGC TGCGTGAACTTCGACTACTCCCGCACCCGCCGCTGCGTGGGCCGCCGCGACCTGCGCCCCGCCAACACCACCTCCACCTGGGAG CCCCCCGTGTCCTCCGACGACGAGGCCTCCTCCCAGTCCAAGCCCCTGGCCACCCAGCCCCCCGTGCTGGCCCTGTCCAACGCC CCCCCCCGCCGCGTGTCCCCCACCCGCGGCCGCCGCCGCCACACCCGCCTGCGCCGCAACtaataaactagtAGTGACTGACTAGGA TCTGGTTACCACTAAACCAGCCTCAAGAACACCCGAATGGAGTCTCTAAGCTACATAATACCAACTTACACTTACAAAATGTTGT CCCCCAAAATGTAGCCATTCGTATCTGCTCCTAATAAAAAGAAAGTTTCTTCACATTCTAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 190).

[0239] In other embodiments, a polyribonucleotide encoding an exemplary polypeptide comprising an HSV-2 gL glycoprotein or immunogenic fragment thereof, comprises or consists of the following nucleotide sequence: GGCTCCCAGGCCACCGAGTACGTGCTGCGCTCCGTGATCGCCAAGGAGGTGGGCGACATCCTGCGCGTGCCCTGCATGCGCACCCC CGCCGACGACGTGTCCTGGCGCTACGAGGCCCCCTCCGTGATCGACTACGCCCGCATCGACGGCATCTTCCTGCGCTACCACTGCC CCGGCCTGGACACCTTCCTGTGGGACCGCCACGCCCAGCGCGCCTACCTGGTGAACCCCTTCCTGTTCGCCGCCGGCTTCCTGGAG GACCTGTCCCACTCCGTGTTCCCCGCCGACACCCAGGAGACCACCACCCGCCGCGCCCTGTACAAGGAGATCCGCGACGCCCTGGG CTCCCGCAAGCAGGCCGTGTCCCACGCCCCCGTGCGCGCCGGCTGCGTGAACTTCGACTACTCCCGCACCCGCCGCTGCGTGGGCC GCCGCGACCTGCGCCCCGCCAACACCACCTCCACCTGGGAGCCCCCCGTGTCCTCCGACGACGAGGCCTCCTCCCAGTCCAAGCCCC TGGCCACCCAGCCCCCCGTGCTGGCCCTGTCCAACGCCCCCCCCCGCCGCGTGTCCCCCACCCGCGGCCGCCGCCGCCACACCCGCPage 49 of 182 12197519v1CTGCGCCGCAAC (SEQ ID NO: 191).

[0240] In some embodiments, a polyribonucleotide encoding an exemplary polypeptide comprising a gL glycoprotein or an immunogenic fragment thereof, comprises a homology greater than 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 191 or SEQ ID NO: 191. HSV Glycoprotein I

[0241] In some embodiments, compositions of the present disclosure further comprise an RNA or a nucleoside- modified RNA encoding HSV glycoprotein I (gI), or an immunogenic fragment thereof. In some embodiments, an RNA encoding an HSV gI, or an immunogenic fragment thereof, comprises an RNA encoding HSV-1 gI, or an immunogenic fragment thereof. In some embodiments, an RNA encoding an HSV gI, or an immunogenic fragment thereof, comprises an RNA encoding HSV-2 gI, or an immunogenic fragment thereof.

[0242] In some embodiments, disclosed herein is a single composition comprising an RNA encoding an HSV gB and an RNA encoding an HSV gI, or immunogenic fragments thereof. In some embodiments, said gB and said gI RNAs are part of a single polyribonucleotide. In some embodiments, disclosed herein are two individual compositions to be administered as part of a single treatment, the first composition comprising an RNA encoding an HSV gB or an immunogenic fragment thereof, and the second composition comprising an RNA encoding an HSV GI or immunogenic fragment thereof.

[0243] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, some uridine residues are 1-methyl-pseudouridine. In another embodiment, the nucleotide sequence of the RNA encoding an HSV-1 or HSV-2 gI fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof.

[0244] In some embodiments, a polyribonucleotide encoding an exemplary polypeptide comprising an HSV-2 gI glycoprotein or an immunogenic fragment thereof, comprises or consists of SEQ ID NO: 184.

[0245] In some embodiments, the present disclosure provides a polyribonucleotide encoding an HSV-2 gB glycoprotein or immunogenic fragment thereof, or a truncated HSV-2 gI glycoprotein or immunogenic fragment thereof. In some embodiments, the present disclosure provides a polyribonucleotide encoding the full-length or non- truncated HSV-2 gI glycoprotein. HSV-2 gI glycoprotein is also termed herein gI2, having all the same features and limitations.

[0246] In some embodiments, a polyribonucleotide utilized in the methods, compositions, and combinations of the present disclosure encodes a full-length HSV-2 gI glycoprotein comprising SEQ ID NO: 30.

[0247] In some embodiments, a polyribonucleotide utilized in the methods, compositions, and combinations of the present disclosure encodes an HSV-2 gI immunogenic fragment, comprising amino acids 25-230 of HSV-2 gI of SEQ ID NO: 30.

[0248] In some embodiments, an HSV-2 gI glycoprotein or immunogenic fragment thereof, or truncated HSV-2 gI glycoprotein or immunogenic fragment thereof, comprises or consists of an amino acid sequence comprising a homology greater than 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 30.Page 50 of 182 12197519v1

[0249] In some embodiments, a polyribonucleotide utilized in the methods, compositions, and combinations of the present disclosure further encodes a signal sequence encoding a signal peptide. In some embodiments, the signal peptide is an IL2 signal peptide (SEQ ID NO: 30).

[0250] In some embodiments, an HSV-2 gI glycoprotein or immunogenic fragment thereof, or truncated HSV-2 gI glycoprotein or immunogenic fragment thereof, encoded by modified RNA utilized in the methods, compositions, and combinations of the present disclosure comprises or consists of the amino acid sequences as set forth in any of the following GenBank Accession Numbers: ABW83313.1, ABW83327.1, ABW83385.1, ABW83397.1, AHG54731.1, AKC42829.1, AKC59306.1, AKC59377.1, AKC59519.1, AKC59590.1, AQZ56891.1, AQZ56962.2, AQZ58027.1, ATD86571.1, ATD86726.1, BAA00021.1, QAU10475.1, QAU10768.1, QBC74573.1, QBH76746.1, QBH76909.1, QBH78328.1, QBH78911.1, QBH80193.1, QBH80706.1, QBH80794.1, QBH81814.1, QBH82569.1, QBH83266.1, QBH84231.1, QBH85083.1, QBH85656.1, or SPT06174.1.

[0251] In some embodiments, a polyribonucleotide for use in the methods, compositions, and combinations of the present disclosure encodes, inter alia, an HSV-2 gI glycoprotein or immunogenic fragment thereof, or truncated HSV-2 gI glycoprotein or immunogenic fragment thereof, (e.g., amino acids 25-230) comprising the following nucleotide sequence: GCCCAGGAGUCCUGGGCCGGCCCCACCGUGUCCCUGGUGUCCGACUCCCUGGUGGACGCCGGCGCCGUGGGCCCCCAGGGCU UCGUGGAGGAGGACCUGCGCGUGUUCGGCGAGCUGCACUUCGUGGGCGCCCAGGUGCCCCACACCAACUACUACGACGGCAU CAUCGAGCUGUUCCACUACCCCCUGGGCAACCACUGCCCCCGCGUGGUGCACGUGGUGACCCUGACCGCCUGCCCCCGCCGCC CCGCCGUGGCCUUCACCCUGUGCCGCUCCACCCACCACGCCCACUCCCCCGCCUACCCCACCCUGGAGCUGGGCCUGGCCCGC CAGCCCCUGCUGCGCGUGCGCACCGCCACCCGCGACUACGCCGGCCUGUACGUGCUGCGCGUGUGGGUGGGCUCCGCCACCA ACGCCUCCCGCUUCGUGCUGGGCGUGGCCCUGUCCGCCAACGGCACCUUCGUGUACAACGGCUCCGACUACGGCUCCUGCGA CCCCGCCCAGCUGCCCUUCUCCGCCCCCCGCCUGGGCCCCUCCUCCGUGUACACCCCCGGCGCCUCCCGCCCCACCCCCCCCC GCACCACCACCCCCCCCUCCUCCCCCCGCGACCCCACCCCCGCCCCCGGCGACACCGGCACCCCCGCCCCCGCCUCCGGCGAGA UCGCCCCCCCCAACUCCACCCGCUCCGCCUCCGAGUCCCGCCACCGCUAA (SEQ ID NO: 192)

[0252] In some embodiments, a nucleotide sequence of the polyribonucleotide encoding an HSV-2 gI glycoprotein or immunogenic fragment thereof, or truncated HSV-2 gI glycoprotein or immunogenic fragment thereof, comprises a homology greater than 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 192.

[0253] In some embodiments, a polyribonucleotide encoding a polypeptide comprising an HSV-2 gI glycoprotein or immunogenic fragment thereof, or truncated HSV-2 gI glycoprotein or immunogenic fragment thereof, as described herein further comprises a signal sequence that encodes a signal peptide. In other embodiments, a polyribonucleotide further comprises nucleotide sequences that are not protein-coding, for e.g., a 5’UTR, a 3’UTR, polyA tail, etc.

[0254] In some embodiments, a polyribonucleotide for use in the methods, compositions, and combinations of the present disclosure comprises or consists of, inter alia, a nucleotide sequence encoding an immunogenic fragment of HSV-2 gI glycoprotein, a signal sequence encoding a signal peptide, and non-protein coding sequences (e.g., a 5’UTR, a 3’UTR, a polyA tail, etc.).Page 51 of 182 12197519v1

[0255] In some embodiments, the polyribonucleotide comprises or consists of the following nucleotide sequence: AGCAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCA UUUCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAACGCCACCAUGGCCAUCUCCGGCGUGCC CGUGCUGGGCUUCUUCAUCAUCGCCGUGCUGAUGUCCGCCCAGGAGUCCUGGGCCGGCCCCACCGUGUCCCUGGUGUCC GACUCCCUGGUGGACGCCGGCGCCGUGGGCCCCCAGGGCUUCGUGGAGGAGGACCUGCGCGUGUUCGGCGAGCUGCACUUCGU GGGCGCCCAGGUGCCCCACACCAACUACUACGACGGCAUCAUCGAGCUGUUCCACUACCCCCUGGGCAACCACUGCCCCCGCGUG GUGCACGUGGUGACCCUGACCGCCUGCCCCCGCCGCCCCGCCGUGGCCUUCACCCUGUGCCGCUCCACCCACCACGCCCACUCCC CCGCCUACCCCACCCUGGAGCUGGGCCUGGCCCGCCAGCCCCUGCUGCGCGUGCGCACCGCCACCCGCGACUACGCCGGCCUGUA CGUGCUGCGCGUGUGGGUGGGCUCCGCCACCAACGCCUCCCGCUUCGUGCUGGGCGUGGCCCUGUCCGCCAACGGCACCUUCGU GUACAACGGCUCCGACUACGGCUCCUGCGACCCCGCCCAGCUGCCCUUCUCCGCCCCCCGCCUGGGCCCCUCCUCCGUGUACACC CCCGGCGCCUCCCGCCCCACCCCCCCCCGCACCACCACCCCCCCCUCCUCCCCCCGCGACCCCACCCCCGCCCCCGGCGACACCGGC ACCCCCGCCCCCGCCUCCGGCGAGAUCGCCCCCCCCAACUCCACCCGCUCCGCCUCCGAGUCCCGCCACCGCUAAACUAGUAGUGA CUGACUAGGAUCUGGUUACCACUAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUAC AAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQID NO: 193)

[0256] In some embodiments, the nucleotide sequence of the modified polyribonucleotide encoding a truncated HSV-2 gI comprises a homology greater than 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 193.

[0257] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 193). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 193). In some embodiments, italicized residues represent 3’ untranslated sequences after which follows the poly adenylation tail (SEQ ID NO: 193).

[0258] In some embodiments, an HSV gI glycoprotein or immunogenic fragment thereof, or truncated HSV gI glycoprotein or immunogenic fragment thereof, encoded by a polyribonucleotide comprises or consists of an HSV-1 gI domain involved in cell-to-cell spread. In another embodiment, an HSV gI glycoprotein or immunogenic fragment thereof, or truncated HSV gI glycoprotein or immunogenic fragment thereof, encoded by a polyribonucleotide utilized in the methods, compositions, and combinations of the present disclosure comprises or consists of an immune evasion domain. In another embodiment, an HSV gI glycoprotein or immunogenic fragment thereof, or truncated HSV gI glycoprotein or immunogenic fragment thereof, encoded by a polyribonucleotide utilized in the methods, compositions, and combinations of the present disclosure comprises or consists of a portion of an immune evasion domain.

[0259] In another embodiment, an HSV gI glycoprotein or immunogenic fragment thereof, or truncated HSV gI glycoprotein or immunogenic fragment thereof, encoded by a polyribonucleotide utilized in the methods, compositions, and combinations of the present disclosure is immunoprotective. A protective immune response generally involves, in some embodiments, an antibody response. In another embodiment, mutants, sequence conservative variants, and functional conservative variants of HSV gI glycoproteins or immunogenic fragments thereof, or truncated HSV gI glycoproteins or immunogenic fragments thereof, are useful in methods, compositions,Page 52 of 182 12197519v1and combinations of the present disclosure, provided that all such variants retain the required immunoprotective effect.

[0260] In some embodiments, an HSV gI glycoprotein or immunogenic fragment thereof, or truncated HSV gI glycoprotein or immunogenic fragment thereof, can be derived from any strain of HSV. In some embodiments, an HSV gI glycoprotein or immunogenic fragment thereof, or truncated HSV gI glycoprotein or immunogenic fragment thereof, can be derived from sequence variants of HSV, as found in HSV-infected individuals.

[0261] In some embodiments, the present disclosure provides an RNA or a nucleoside-modified RNA encoding a polypeptide that comprises the ectodomain of HSV-2 gB, or an immunogenic fragment thereof, wherein the RNA further comprises a secretory signal sequence. In some embodiments, the secretory signal sequence encodes an IL2 secretory signal polypeptide. In some embodiments, the IL2 secretory signal polypeptide comprises or consists of the amino acid sequence as set forth in SEQ ID NO: 2. In another embodiment, the IL2 secretory signal polypeptide is encoded by the ribonucleic acid sequence as set forth in SEQ ID NO: 3.

[0262] In some embodiments, a single polyribonucleotide encodes both the HSV gB glycoprotein or immunogenic fragment thereof, and the additional HSV glycoprotein or immunogenic fragment thereof. In other embodiments, the HSV gB glycoprotein or immunogenic fragment thereof, and the additional HSV glycoprotein or immunogenic fragment thereof are encoded by separate polyribonucleotides. In some embodiments, a single polyribonucleotide encodes both a truncated HSV gB glycoprotein or immunogenic fragment thereof, and a truncated additional HSV glycoprotein or immunogenic fragment thereof. In other embodiments, the truncated HSV gB glycoprotein or immunogenic fragment thereof, and an additional truncated HSV glycoprotein or immunogenic fragment thereof are encoded by separate polyribonucleotides.

[0263] In some embodiments, a combination of the present disclosure comprises a polyribonucleotide encoding an immunogenic fragment of a Herpes Simplex Virus (HSV)-2 glycoprotein B (gB) antigen, an immunogenic fragment of an HSV-2 glycoprotein C (gC) antigen, an immunogenic fragment of an HSV-2 glycoprotein D (gD) antigen, and an immunogenic fragment of an HSV-2 glycoprotein E (gE) antigen. In other embodiments, a combination of the present disclosure comprises a polyribonucleotide encoding an immunogenic fragment of an HSV-2 gB antigen, an immunogenic fragment of an HSV-2 gC antigen, an immunogenic fragment of an HSV-2 gD antigen, an immunogenic fragment of an HSV-2 gE antigen, and an immunogenic fragment of an HSV-2 gI antigen.

[0264] In other embodiments, a combination of the present disclosure comprises a polyribonucleotide encoding an immunogenic fragment of an HSV gB antigen, an immunogenic fragment of an HSV gC antigen, and an immunogenic fragment of an HSV gD antigen.

[0265] In some embodiments, a polyribonucleotide of the present disclosure encodes an HSV polypeptide, or fragment thereof, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 1.

[0266] In some embodiments, methods of the present disclosure comprise administering to a subject an HSV polypeptide, or fragment thereof, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at leastPage 53 of 182 12197519v193%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 1. TABLE 1: Exemplary amino acid sequences of HSV immunogens Sequence Amino acid sequence SEQ ID NO: Name HSV-1 gB APTSPGTPGVAAATQAANGGPATPAPPPLGAAPTGDPKPKKNKKPKNPTPPRPAG 10Page 54 of 182 12197519v1Sequence Amino acid sequence SEQ ID NO: Name (mutated LKYNPSRVEAFHRYGTTVNCIVEEVDARSVYPYDEFVLATGDFVYMSPFYGYREGSPage 55 of 182 12197519v1Sequence Amino acid sequence SEQ ID NO: Name HSV-2 gB MRGGGLICALVVGALVAAVASAAPAAPAAPRASGGVAATVAANGGPASRPPPVPSP 7Page 56 of 182 12197519v1Sequence Amino acid sequence SEQ ID NO: Name stop codons YDEFVLATGDFVYMSPFYGYREGSHTEHTSYAADRFKQVDGFYARDLTTKARATSPage 57 of 182 12197519v1Sequence Amino acid sequence SEQ ID NO: Name TAPSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGPage 58 of 182 12197519v1Sequence Amino acid sequence SEQ ID NO: Name HSV-2 gD KYALADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITV 36Page 59 of 182 12197519v1Sequence Amino acid sequence SEQ ID NO: Name PPPTPPRRPPVAPPTHPRVIPEVSHVRGVTVHMETPEAILFAPGETFGTNVSIHAIA [ribonucleic acid sequence as set forth in SEQ ID NO: 6, comprising sequences encoding an IL2 signal polypeptide and a truncated HSV-2 glycoprotein B.

[0268] In some embodiments, a polyribonucleotide of the present disclosure comprises a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 2.

[0269] In some embodiments, methods of the present disclosure comprise administering to a subject a polyribonucleotide comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 2.Page 60 of 182 12197519v1TABLE 2: Exemplary nucleic acid sequences of HSV immunogens Sequence Nucleic acid sequence SEQ ID NO: Name HSV-2 gB GCCCCCGCCGCCCCCGCCGCCCCCCGCGCCTCCGGCGGCGTGGCCGCCACCGT 5Page 61 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name AGTACGTGCGCAACAACATGGAGACCACCGCCTTCCACCGCGACGACCACGAGAPage 62 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GTACAGCCTGTCCAGAGTGGACCTGGGCGATTGCATCGGCAGAGATGCCAGAGPage 63 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CGAATGCCTCCGTCGAGCGAATTAAGACCACATCTAGCATCGAATTTGCGCGCCPage 64 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GATTGTGCAGAATTCAATGAGAGTGTCTTCAAGACCTGGAACCTGCTACTCAAGPage 65 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CACCCGGCACGAGATCAAGGATAGCGGCCTGCTGGATTACACCGAGGTGCAGAPage 66 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GTGCTGCAACTTCAGCGAAATCCAATGAAGGCGCTCTACCCTCTCACAACAAAAPage 67 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name AGCAAGATACTCTCCTCTGCACAATGAAGATGAAGCTGGAGATGAAGATGAACTPage 68 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GCCAAGGGTGTGTGTAGATCCACTGCTAAATACGTCAGGAACAATATGGAAACAPage 69 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CCTCAGTACTACCTGGCAACAGGAGGATTTCTGATTGCTTACCAGCCTCTGCTGPage 70 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CGTCACCACCGTCAGCACCTTCATCGACCTGAACATCACCATGCTGGAGGACCAPage 71 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name ACATCGCTCCCTATAAATTCAAGGCCACCATGTACTATAAGGACGTTACAGTCTCPage 72 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name TGGATCTGGGAGATTGCATTGGGAGAGATGCAAGAGAAGCAATTGATAGAATGPage 73 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GACGCGCTCGAGCCGTGCACCGTGGGCCACCGGCGCTACTTCATCTTCGGCGGPage 74 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CACCGTGTCCACCGTGACCTCCGCCGCCGTGGGCGGCCAGGGCCCCCCCCGCACPage 75 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name HSV-2 gC TCTCCCGGCAGAACCATCACAGTGGGCCCTAGAGGCAACGCCTCTAATGCCGCT 53Page 76 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GACACAGGGGATGTACTACTGGGTGTGGGGCCGTACTGACCGCCCTTCCGCATPage 77 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CGAGCTGCGGTCGACCAGGAACAGCTACCATTCGCAGCACTCTGCCCGTGTCCTPage 78 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CAGAGATTGGTACCGCCCCCAGTTTGGAGGAAGTGATGGTCAACGTGTCCGCACPage 79 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CTGTCAGCTGACCTGGCACAGAGACTCCGTAAGCTTCAGCCGTAGAAACGCCTCPage 80 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name (28-426) AGCCAAGAAAAGCAACCAAATCCAAAGCATCCACAGCAAAACCTGCACCTCCTCCPage 81 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CCATGCTGTGCTGGAAGGACAGCCGTTCAAGGCAACATGCACAGCAGCCACTTAPage 82 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GAACAGACTGAGTATATTTGCAGACTGGCTGGATACCCGGATGGGATTCCTGTCPage 83 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CATCGGCATGCTGCCCCGCTTCATCCCCGAGAACCAGCGCACCGTGGCCCTGTAPage 84 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CCCTGAAGATCGCAGGATGGCATGGGCCCAAACCACCTTATACCTCTACGTTGCPage 85 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GTGACACCACTAATGCGACACAGCCAGAACTTGTGCCTGAGGATCCTGAAGATAPage 86 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name AGTGCCTACACCTACACCTGACGACTACGACGAGGAAGATGACGCTGGCGTCAGPage 87 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CGTCGTGGAGCAGCACCTCCCCCAACGGCAGCCAGAACCAGTGGAGCCCACTCGPage 88 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GAAAGAACCCCCGTGTCCGTGCCCCCTCCCACCCCTCCCCGCAGACCCCCTGTGPage 89 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name TGTGGTCGAGCAGCACTTGCCCCAGCGACAACCCGAACCAGTGGAGCCAACCAGPage 90 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GCCTCCGTGCTGCCCCGCCCCACCATCACCATGGAGTTCACCGGCGACCACGCCPage 91 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CACATTCTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAPage 92 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name stop codons CGCTCCTCCACCTCTTGGAGCTGCTCCTACAGGCGACCCCAAGCCTAAGAAGAAPage 93 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name TGAGGGGATAGCTGTCGTATTTAAAGAAAATATTGCTCCTTATAAATTCAAGGCPage 94 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name AAACCTGCAAATGCTGCAACCAGAACCTCAAGAGGATGGCACACCACAGATCTGPage 95 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GTGGCCTGGGACTGGGTGCCAAAGCGCCCGTCGGTCTGCACCATGACCAAGTGPage 96 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CGAGCTGTATGTGCGCGAGCATCTGAGGGAACAGAGCCGGAAGCCTCCTAATCCPage 97 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GAGGATCAGCTCTCGTCCCGGAGCCTGCTATAGCAGACCACTGGTGTCCTTCCGPage 98 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name TGCTGATATTGATACAGTGATCCATGCTGATGCAAATGCTGCAATGTTTGCTGGPage 99 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name gB1 full seq ATGCATCAGGGCGCTCCATCTTGGGGTAGACGTTGGTTCGTTGTGTGGGCCCT 212Page 100 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GAACAACCTGGAGACTACCGCATTCCATCGTGACGATCACGAGACTGATATGGAPage 101 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name ACATCAAAGTGGGACAGCCTCAGTACTACCAGGCAAATGGAGGATTTCTGATTGPage 102 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GCCACGAGATCAAGGACAGCGGCCTGCTGGACTACACGGAGGTCCAGCGCCGCPage 103 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name TCCCATTCGAAGAAGTGATTGACAAGATCAACGCCAAAGGCGTGTGCAGATCAAPage 104 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GAGGATTTCTGATTGCTTACCAGCCTCTGCTGTCAAATACCCTGGCTGAACTGTPage 105 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name CCCAGCCCCGCTGGTCCTACTACGACTCCTTCTCCGCCGTGTCCGAGGACAACCPage 106 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GCGCATGGAGACCCCCGAGGCCATCCTGTTCTCCCCCGGCGAGGCCTTCTCCACPage 107 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GACCGACTCCGGCCTGTACACCCTGTCCGTGGTGGGCCTGTCCGACGAGGCCCPage 108 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name Truncated GGCTCCCAGGCCACCGAGTACGTGCTGCGCTCCGTGATCGCCAAGGAGGTGGG 185Page 109 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name GCCGACTCCGTGTTCCTGAACGTGTCCGTGCTGGACCCCGCCGCCCGCCTGCGCPage 110 of 182 12197519v1Sequence Nucleic acid sequence SEQ ID NO: Name ACCAGAAGAACCTGTTCTGGCTGCCCGACCACTTCTCCCCCTGCGCCGCCTCCC Secretory Signal

[0270] In some embodiments, a polyribonucleotide or RNA disclosed herein further comprises a secretory signal sequence encoding a secretory signal polypeptide, e.g., that is functional in mammalian cells. In some embodiments, a secretory signal sequence encodes a modified secretory signal polypeptide (e.g., comprising amino acid substitutions or amino acid additions). In some embodiments, a secretory signal sequence is a codon optimized secretory signal sequence.

[0271] In some embodiments, a utilized secretory signal sequence is a heterologous secretory signal sequence. In some embodiments, a heterologous secretory signal sequence comprises or consists of a non-human secretory signal sequence. In some embodiments, a heterologous secretory signal sequence comprises or consists of a viral secretory signal sequence. In some embodiments, a viral secretory signal sequence comprises or consists of an HSV secretory signal sequence (e.g., an HSV-1 or HSV-2 secretory signal sequence). In some embodiments, a secretory signal sequence comprises or consists of an HSV-1 secretory signal sequence. In some embodiments, a secretory signal sequence comprises or consists of an HSV-2 secretory signal sequence. In some embodiments, a secretory signal sequence encodes a secretory signal polypeptide characterized by a length of about 15 to 30 amino acids. In some embodiments, a secretory signal sequence encodes a secretory signal polypeptide that preferably allows transport of an HSV-1 glycoprotein, or immunogenic fragment thereof, an HSV-2 glycoprotein, or immunogenic fragment thereof, or both, with which it is associated into a defined cellular compartment, preferably a cell surface, endoplasmic reticulum (ER) or endosomal-lysosomal compartment.

[0272] In some embodiments, a secretory signal sequence is the native secretory signal sequence of the encoded glycoprotein.

[0273] Suitable secretory signal sequences are generally known in the art; for example, the signal sequence of human tissue plasminogen activator (tPA). In general, signal sequences encode signal peptides that are short peptides located at the N-terminus of proteins, carrying information for protein secretion. In some embodiments, thePage 111 of 182 12197519v1signal peptide may be from a prokaryote or from a eukaryote. In some embodiments, the signal peptide comprises 25–30 residues. In other embodiments, longer signal peptides, such eukaryotic signal sequences may be utilized. In some embodiments, the signal peptide comprises up to about 140 residues. In some embodiments, the secretory signal sequence in polyribonucleotides described herein are HSV secretory signals.

[0274] In some embodiments, the secretory signal sequence encodes an IL2 secretory signal polypeptide. In some embodiments, the IL2 secretory signal polypeptide comprises or consists of the amino acid sequence as set forth in SEQ ID NO: 2. In another embodiment, the IL2 secretory signal polypeptide is encoded by the ribonucleic acid sequence as set forth in SEQ ID NO: 3. In another embodiment, the secretory signal sequence encodes an HSV gB secretory signal polypeptide. In some embodiments, the HSV gB secretory signal polypeptide comprises or consists of the amino acid sequence as set forth in SEQ ID NO: 4. In another embodiment, the secretory signal sequence encodes an HSV gD secretory signal polypeptide, which, in one embodiment, is an HSV-2 gD secretory signal polypeptide. In another embodiment, the secretory signal sequence encodes an HSV protein secretory signal polypeptide, which in one embodiment, comprises an HSV glycoprotein secretory signal. In one embodiment, the HSV glycoprotein secretory signal is an HSV-2 glycoprotein secretory signal.

[0275] In some embodiments, the present disclosure provides a polyribonucleotide encoding a polypeptide that comprises the ectodomain of HSV-2 gB, or an immunogenic fragment thereof, wherein the RNA further comprises a secretory signal sequence encoding an HSV gB secretory signal polypeptide. In some embodiments, the HSV gB secretory signal polypeptide comprises or consists of the amino acid sequence as set forth in SEQ ID NO: 4.

[0276] In some embodiments, an RNA as disclosed herein comprises a secretory signal sequence encoding a secretory signal polypeptide listed in Table 3, or a secretory signal peptide having 1, 2, 3, 4, or 5 amino acid differences thereto. In some embodiments, a secretory signal peptide is selected from those listed in Table 3 and functionally connected to the N-terminus of an HSV immunogen selected from those listed in Table 1. TABLE 3: Exemplary signal peptides Sequence Name Amino Acid Sequence SEQ ID NO:Page 112 of 182 12197519v1Sequence Name Amino Acid Sequence SEQ ID NO: HSV-1 gD Signal Sequence MGGAAARLGAVILFVVIVGLHGVRG 97 [ nalsequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence listed in Table 4. In some embodiments, a secretory signal sequence is selected from those listed in Table 4 and functionally connected (i.e., in frame) to the 5' end of an HSV immunogen nucleic acid sequence selected from those listed in Table 2. TABLE 4: Exemplary signal sequences Sequence Name Version RNA Sequence SEQ ID NO:Page 113 of 182 12197519v1Sequence Name Version RNA Sequence SEQ ID NO: HSV-1 gB Version 3 ATGCACCAGGGAGCACCTTCTTGGGGAAGAAGATGGTTTG 119Page 114 of 182 12197519v1Sequence Name Version RNA Sequence SEQ ID NO: HSV-2 gB AP Version 1 ATGAGAGGCGGCGGACTGATTTGTGCCCTGGTTGTGGGAG 232Page 115 of 182 12197519v1Sequence Name Version RNA Sequence SEQ ID NO: HSV-2 gD KYAL Version 3 ATGGGCAGACTGACCTCCGGCGTGGGCACCGCCGCCCTGC 134Page 116 of 182 12197519v1Sequence Name Version RNA Sequence SEQ ID NO: HSV-2 gC Version 2.1 ATGGCACTAGGGAGAGTGGGATTAGCTGTGGGTCTGTGGG 151Page 117 of 182 12197519v1Sequence Name Version RNA Sequence SEQ ID NO: HSV-2 gI (1-18) Version 2 ATGCCCGGCAGAAGCCTCCAGGGACTGGCTATCCTGGGGC 172Exemplary Nucleotide Sequence Features

[0278] In some embodiments, the polyribonucleotide or RNA disclosed herein further comprises a 5′ untranslated region. In some embodiments, the polyribonucleotide or RNA disclosed herein further comprises a 3′ untranslated region. In some embodiments, nucleotide sequences described herein may comprise a 5’ cap, which may be incorporated during transcription, or joined to a nucleotide sequence post-transcription.

[0279] In some embodiments, the polyribonucleotide or RNA disclosed herein further comprises a poly-A tail. In some embodiments, the present disclosure provides an RNA construct comprising (i) a 5′ untranslated region, (ii) a polyribonucleotide as disclosed herein, (iii) a 3′ untranslated region, and (iv) a poly-A tail sequence. In some embodiments, the 5’ untranslated region is from the tobacco etch virus. In some embodiments, the 3’ untranslated region is from Xenopus beta globin.

[0280] In some embodiments, the polyribonucleotide or RNA disclosed herein further comprises a cap- independent translational enhancer. In some embodiments, the cap-independent translational enhancer is a tobacco etch virus (TEV) cap-independent translational enhancer. In another embodiment, the cap-independent translational enhancer is any other cap-independent translational enhancer known in the art. Each possibility represents a separate embodiment of the present disclosure.Page 118 of 182 12197519v1

[0281] In other embodiments, polyribonucleotides of the methods, compositions, and combinations of the present disclosure further comprise an internal ribosome entry site (IRES). In some embodiments, the IRES comprises a 5'Leader IRES and intercistronic IRES in the 1.8-kb family of immediate early transcripts (IRES)1; Aphthovirus IRES; Cripavirus IRES; Hepatitis A IRES; Hepatitis C IRES; Kaposi's sarcoma-associated herpesvirus IRES; Pestivirus IRES; Picornavirus IRES; Rhopalosiphum padi virus IRES; or a combination thereof.

[0282] In other embodiments, polyribonucleotides of the methods, compositions, and combinations of the present disclosure further comprise a cap-independent translational enhancer (CITE). In another embodiment, polyribonucleotides of the methods, compositions, and combinations of the present disclosure do not comprise a CITE. In some embodiments, the CITE is a tobacco etch virus (TEV) cap-independent translational enhancer. In other embodiments, the CITE comprises TED (translation enhancer domain); BTE (BYDV-like translation element); PTE (PMV-like translation element); TSS (T-shaped structure); Y-shaped; I-shaped; or a combination thereof. In other embodiments, the cap-independent translational enhancer is any other cap-independent translational enhancer known in the art. Each possibility represents a separate embodiment of the present disclosure.

[0283] In some embodiments, the polyribonucleotide or RNA disclosed herein further comprises an m7GpppG cap, 3′-O-methyl-m7GpppG cap, or anti-reverse cap analog.

[0284] In some embodiments, the polyribonucleotide or RNA disclosed herein further comprises one or more non-protein-coding sequences. In some embodiments, the non-protein-coding sequence performs one or more of the following: increases protein expression, modulates innate and adaptive immunogenicity, improves delivery and localization within the cell. In some embodiments, the non-protein-coding sequence is involved in pre-mRNA processing, RNA stability, and translation initiation. Non-protein-coding sequences having the above functions are generally known in the art.

[0285] In some embodiments, a nucleotide sequence provided herein encodes one or more glycoproteins (e.g., HSV gB), or an immunogenic fragment thereof. In some embodiments, a polyribonucleotide comprises a 5’ cap, a 5’UTR, a nucleotide sequence that encodes one or more glycoproteins (e.g., HSV gB), or an immunogenic fragment thereof, a 3’ UTR, and a polyA tail. 1. 5' Cap

[0286] A structural feature of natural messenger RNA (mRNA) is a cap structure at the five-prime end (5'). Natural eukaryotic mRNA comprise a 7-methylguanosine cap linked to the mRNA via a 5´ to 5´-triphosphate bridge resulting in a cap0 structure (m7GpppN). Further modifications can occur at the 2'-hydroxyl-group (2’-OH) (e.g., the 2'-hydroxyl group may be methylated to form 2'-O-Me) of the first and subsequent nucleotides producing “cap1” and “cap2” five-prime ends, respectively). Diamond, et al., (2014) Cytokine & growth Factor Reviews, 25:543–550, which is incorporated herein by reference in its entirety, reported that cap0-RNA cannot be translated as efficiently as cap1- RNA in which the role of 2'-O-Me in the penultimate position at the RNA 5’ end is determinant. Lack of the 2'-O-met has been shown to trigger innate immunity and activate an interferon (IFN) response. Daffis, et al. (2010) Nature, 468:452-456; and Züst et al. (2011) Nature Immunology, 12:137-143, each of which is incorporated herein by reference in its entirety.Page 119 of 182 12197519v1

[0287] RNA capping is well researched and is described, e.g., in Decroly E et al. (2012) Nature Reviews 10: 51- 65; and in Ramanathan A. et al., (2016) Nucleic Acids Res; 44(16): 7511–7526, the entire contents of each of which are hereby incorporated by reference. For example, in some embodiments, a 5’-cap structure which may be suitable in the context of the present invention is a cap0 (methylation of the first nucleobase, e.g., m7GpppN), cap1 (additional methylation of the ribose of the adjacent nucleotide of m7GpppN), cap2 (additional methylation of the ribose of the 2nd nucleotide downstream of the m7GpppN), cap3 (additional methylation of the ribose of the 3rd nucleotide downstream of the m7GpppN), cap4 (additional methylation of the ribose of the 4th nucleotide downstream of the m7GpppN), ARCA (“anti-reverse cap analogue”), modified ARCA (e.g. phosphothioate modified ARCA), inosine, N1 -methyl-guanosine, 2’-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino- guanosine, LNA-guanosine, and 2-azido-guanosine.

[0288] The term “5'-cap” as used herein refers to a structure found on the 5'-end of an RNA (e.g., mRNA) and generally includes a guanosine nucleotide connected to an RNA (e.g., mRNA) via a 5'- to 5'-triphosphate linkage (also referred to as Gppp or G(5')ppp(5')). In some embodiments, a guanosine nucleoside included in a 5’ cap may be modified, for example, by methylation at one or more positions (e.g., at the 7-position) on a base (guanine), and / or by methylation at one or more positions of a ribose. In some embodiments, a guanosine nucleoside included in a 5’ cap comprises a 3’O methylation at a ribose (3’OMeG). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine (m7G). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7'-position of guanine and a 3' O methylation at a ribose (m7(3'OMeG)). It will be understood that the notation used in the above paragraph, e.g., “(m27,3'-O)G” or “m7(3'OMeG)”, applies to other structures described herein.

[0289] In some embodiments, providing an RNA with a 5'-cap disclosed herein may be achieved by in vitro transcription, in which a 5'-cap is co-transcriptionally incorporated into an RNA strand, or may be attached to an RNA post-transcriptionally using capping enzymes. In some embodiments, co-transcriptional capping with a cap disclosed herein improves the capping efficiency of an RNA compared to co-transcriptional capping with an appropriate reference comparator. In some embodiments, improving capping efficiency can increase a translation efficiency and / or translation rate of an RNA, and / or increase expression of an encoded protein. In some embodiments, alterations to polynucleotides generate a non-hydrolyzable cap structure which can, for example, prevent decapping and increase RNA half-life.

[0290] In some embodiments, a utilized 5' cap is a cap0, a cap1, or cap2 structure. See, e.g., Fig. 1 of Ramanathan A et al., and Fig. 1 of Decroly E et al., each of which is incorporated herein by reference in its entirety. In some embodiments, an RNA described herein comprises a cap1 structure. In some embodiments, an RNA described herein comprises a cap2 structure.

[0291] In some embodiments, an RNA described herein comprises a cap0 structure. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G). In some embodiments, such a cap0 structure is connected to an RNA via a 5'- to 5'-triphosphate linkage and is also referred to herein as (m7)Gppp. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 2'-position of the ribose of guanosine. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 3'-position of the ribose of guanosine. In some embodiments, a guanosine nucleoside included in aPage 120 of 182 12197519v15' cap comprises methylation at the 7-position of guanine and at the 2'-position of the ribose ((m27,2'-O)G). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine and at the 2'-position of the ribose ((m27,3'-O)G).

[0292] In some embodiments, a cap1 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and optionally methylated at the 2' or 3' position of the ribose, and a 2'O methylated first nucleotide in an RNA ((m2'-O)N1). In some embodiments, a cap1 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and the 3' position of the ribose, and a 2'O methylated first nucleotide in an RNA ((m2'-O)N1). In some embodiments, a cap1 structure is connected to an RNA via a 5'- to 5'-triphosphate linkage and is also referred to herein as, e.g., ((m7)Gppp(2'-O)N1) or (m27,3’-O)Gppp(2'-O)N1), wherein N1 is as defined and described herein. In some embodiments, a cap1 structure comprises a second nucleotide, N2, which is at position 2 and is chosen from A, G, C, or U, e.g., (m7)Gppp(2'-O)N1pN2 or (m27,3’-O)Gppp(2'-O)N1pN2 , wherein each of N1 and N2 is as defined and described herein.

[0293] In some embodiments, a cap2 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and optionally methylated at the 2' or 3' position of the ribose, and 2'O methylated first and second nucleotides in an RNA ((m2’-O)N1p(m2’-O)N2). In some embodiments, a cap2 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and the 3' position of the ribose, and 2'O methylated first and second nucleotides in an RNA. In some embodiments, a cap2 structure is connected to an RNA via a 5'- to 5'- triphosphate linkage and is also referred to herein as, e.g., ((m7)Gppp(2'-O)N1p(2'-O)N2) or (m27,3’-O)Gppp(2'- O)N1p(2'-O)N2), wherein each of N1 and N2 is as defined and described herein.

[0294] In some embodiments, the 5' cap is a dinucleotide cap structure. In some embodiments, the 5' cap is a dinucleotide cap structure comprising N1, wherein N1 is as defined and described herein. In some embodiments, the 5' cap is a dinucleotide cap G*N1, wherein N1 is as defined above and herein, and G* comprises a structure of formula (I): R2 R3 or a salt thereof,wherein each R2 and R3 is -OH or -OCH3; and X is O or S.

[0295] In some embodiments, R2 is -OH. In some embodiments, R2 is -OCH3. In some embodiments, R3 is -OH. In some embodiments, R3 is -OCH3. In some embodiments, R2 is -OH and R3 is -OH. In some embodiments, R2 is - OH and R3 is -CH3. In some embodiments, R2 is -CH3 and R3 is -OH. In some embodiments, R2 is -CH3 and R3 is - CH3.Page 121 of 182 12197519v1

[0296] In some embodiments, X is O. In some embodiments, X is S.

[0297] In some embodiments, the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’-O)GppSpN1), wherein N1 is as defined and described herein. In some embodiments, the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’- O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’-O)GppSpN1), wherein N1 is G. In some embodiments, the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’- O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’-O)GppSpN1), wherein N1 is A, U, or C. In some embodiments, the 5' cap is a dinucleotide cap1 structure (e.g., (m7)Gppp(m2’-O)N1, (m27,2’-O)Gppp(m2’-O)N1, (m27,3’-O)Gppp(m2’-O)N1, (m7)GppSp(m2’-O)N1, (m27,2’-O)GppSp(m2’-O)N1, or (m27,3’-O)GppSp(m2’-O)N1), wherein N1 is as defined and described herein. In some embodiments, the 5' cap is selected from the group consisting of (m7)GpppG (“Ecap0”), (m7)Gppp(m2’-O)G (“Ecap1”), (m27,3’-O)GpppG (“ARCA” or “D1”), and (m27,2’- O)GppSpG (“beta-S-ARCA”). In some embodiments, the 5' cap is (m7)GpppG (“Ecap0”), having a structure of formula (II): or a salt thereof.

[0298] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)G (“Ecap1”), having a structure of formula (III): or a salt thereof.

[0299] In some embodiments, the 5' cap is (m27,3’-O)GpppG (“ARCA” or “D1”), having a structure of formula (IV):Page 122 of 182 12197519v1or a salt thereof

[0300] In some embodiments, the 5' cap is (m27,2’-O)GppSpG (“beta-S-ARCA”), having a structure of formula (V): or a salt

[0301] In some embodiments, the 5' cap is a trinucleotide cap structure. In some embodiments, the 5' cap is a trinucleotide cap structure comprising N1pN2, wherein N1 and N2 are as defined and described herein. In some embodiments, the 5' cap is a dinucleotide cap G*N1pN2, wherein N1 and N2 are as defined above and herein, and G* comprises a structure of formula (VI): R2 R3 or a salt thereof, wherein R2,Page 123 of 182 12197519v1

[0302] In some embodiments, the 5' cap is a trinucleotide cap0 structure (e.g., (m7)GpppN1pN2, (m27,2’- O)GpppN1pN2, or (m27,3’-O)GpppN1pN2), wherein N1 and N2 are as defined and described herein). In some embodiments, the 5' cap is a trinucleotide cap1 structure (e.g., (m7)Gppp(m2’-O)N1pN2, (m27,2’-O)Gppp(m2’- O)N1pN2, (m27,3’-O)Gppp(m2’-O)N1pN2), wherein N1 and N2 are as defined and described herein. In some embodiments, the 5' cap is a trinucleotide cap2 structure (e.g., (m7)Gppp(m2’-O)N1p(m2’-O)N2, (m27,2’- O)Gppp(m2’-O)N1p(m2’-O)N2, (m27,3’-O)Gppp(m2’-O)N1p(m2’-O)N2), wherein N1 and N2 are as defined and described herein. In some embodiments, the 5' cap is selected from the group consisting of (m27,3’-O)Gppp(m2’- O)ApG (“CleanCap AG”, “CC413”), (m27,3’-O)Gppp(m2’-O)GpG (“CleanCap GG”), (m7)Gppp(m2’-O)ApG, (m7)Gppp(m2’-O)GpG, (m27,3’-O)Gppp(m26,2’-O)ApG, and (m7)Gppp(m2’-O)ApU.

[0303] In some embodiments, the 5' cap is (m27,3’-O)Gppp(m2’-O)ApG (“CleanCap AG”, “CC413”), having a structure of formula (VII): or a salt

[0304] In some embodiments, the 5' cap is (m27,3’-O)Gppp(m2’-O)GpG (“CleanCap GG”), having a structure of formula (VIII):Page 124 of 182 12197519v1or a salt there

[0305] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)ApG, having a structure of formula (IX): or a salt

[0306] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)GpG, having a structure of formula (X):Page 125 of 182 12197519v1or a salt the

[0307] In some embodiments, the 5' cap is (m27,3’-O)Gppp(m26,2’-O)ApG, having a structure of formula (XI): or a salt

[0308] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)ApU, having a structure of formula (XII):Page 126 of 182 12197519v1or a salt thereof.

[0309] In some embodiments, the 5' cap is a tetranucleotide cap structure. In some embodiments, the 5' cap is a tetranucleotide cap structure comprising N1pN2pN3, wherein N1, N2, and N3 are as defined and described herein. In some embodiments, the 5' cap is a tetranucleotide cap G*N1pN2pN3, wherein N1, N2, and N3 are as defined above and herein, and G* comprises a structure of formula (XIII): R2 R3 or a salt thereof, wherein R2,

[0310] In some embodiments, the 5' cap is a tetranucleotide cap0 structure (e.g. (m7)GpppN1pN2pN3, (m27,2’-O)GpppN1pN2pN3, or (m27,3’-O)GpppN1N2pN3), wherein N1, N2, and N3are as defined and described herein). In some embodiments, the 5’ cap is a tetranucleotide Cap1 structure (e.g., (m7)Gppp(m2’-O)N1pN2pN3, (m27,2’-O)Gppp(m2’-O)N1pN2pN3, (m27,3’-O)Gppp(m2’-O)N1pN2N3), wherein N1, N2, and N3are as defined and described herein. In some embodiments, the 5' cap is a tetranucleotide Cap2 structure (e.g., (m7)Gppp(m2’-O)N1p(m2’-O)N2pN3, (m27,2’-O)Gppp(m2’-O)N1p(m2’-O)N2pN3, (m27,3’-O)Gppp(m2’-O)N1p(m2’-O)N2pN3), wherein N1, N2, and N3 are as defined and described herein. In some embodiments, the 5' cap is selected from the group consisting of (m27,3’-O)Gppp(m2’-O)Ap(m2’-O)GpG, (m27,3’-O)Gppp(m2’-O)Gp(m2’-O)GpC, (m7)Gppp(m2’-O)Ap(m2’-O)UpA, and (m7)Gppp(m2’-O)Ap(m2’-O)GpG.Page 127 of 182 12197519v1

[0311] In some embodiments, the 5' cap is (m27,3’-O)Gppp(m2’-O)Ap(m2’-O)GpG, having a structure of formula (XIV): or a salt thereof.

[0312] In some embodiments, the 5' cap is (m27,3’-O)Gppp(m2’-O)Gp(m2’-O)GpC, having a structure of formula (XV):Page 128 of 182 12197519v1or a salt thereof.

[0313] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)Ap(m2’-O)UpA, having a structure of formula (XVI): or a salt

[0314] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)Ap(m2’-O)GpG, having a structure of formula (XVII):Page 129 of 182 12197519v1or a salt ther 2. Cap Proximal Sequences

[0315] In some embodiments, a 5' UTR utilized in accordance with the present disclosure comprises a cap proximal sequence, e.g., as disclosed herein. In some embodiments, a cap proximal sequence comprises a sequence adjacent to a 5' cap. In some embodiments, a cap proximal sequence comprises nucleotides in positions +1, +2, +3, +4, and / or +5 of an RNA polynucleotide.

[0316] In some embodiments, a cap structure comprises one or more polynucleotides of a cap proximal sequence. In some embodiments, a cap structure comprises an m7guanosine cap and nucleotide +1 (N1) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7guanosine cap and nucleotide +2 (N2) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7guanosine cap and nucleotides +1 and +2 (N1and N2) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7guanosine cap and nucleotides +1, +2, and +3 (N1, N2, and N3) of an RNA polynucleotide.

[0317] Those skilled in the art, reading the present disclosure, will appreciate that, in some embodiments, one or more residues of a cap proximal sequence (e.g., one or more of residues +1, +2, +3, +4, and / or +5) may be included in an RNA by virtue of having been included in a cap entity (e.g., a cap1 or cap2 structure, etc.); alternatively, in some embodiments, at least some of the residues in a cap proximal sequence may be enzymatically added (e.g., by a polymerase such as a T7 polymerase). For example, in certain exemplified embodiments where a m27,3’-OGppp(m12’-O)ApG cap is utilized, +1 (i.e., N1) and +2 (i.e. N2) are the (m12’-O)A and G residues of the cap, and +3, +4, and +5 are added by a polymerase (e.g., T7 polymerase).Page 130 of 182 12197519v1

[0318] In some embodiments, the 5'’ cap is a dinucleotide cap structure, wherein the cap proximal sequence comprises N1of the 5’ cap, where N1is any nucleotide, e.g., A, C, G or U. In some embodiments, the 5' cap is a trinucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N1 and N2 of the 5' cap, wherein N1 and N2 are independently any nucleotide, e.g., A, C, G or U. In some embodiments, the 5' cap is a tetranucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N1, N2, and N3of the 5' cap, wherein N1, N2, and N3 are any nucleotide, e.g., A, C, G or U.

[0319] In some embodiments, e.g., where the 5' cap is a dinucleotide cap structure, a cap proximal sequence comprises N1 of a 5' cap, and N2, N3, N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and / or +5 of an RNA polynucleotide. In some embodiments, e.g., where the 5' cap is a trinucleotide cap structure, a cap proximal sequence comprises N1 and N2 of a 5' cap, and N3, N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and / or +5 of an RNA polynucleotide. In some embodiments, e.g., where the 5' cap is a tetranucleotide cap structure, a cap proximal sequence comprises N1, N2, and N3 of a the 5’ cap, and N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and / or +5 of an RNA polynucleotide.

[0320] In some embodiments, N1 is A. In some embodiments, N1 is C. In some embodiments, N1 is G. In some embodiments, N1 is U. In some embodiments, N2 is A. In some embodiments, N2 is C. In some embodiments, N2 is G. In some embodiments, N2 is U. In some embodiments, N3 is A. In some embodiments, N3 is C. In some embodiments, N3 is G. In some embodiments, N3 is U. In some embodiments, N4 is A. In some embodiments, N4 is C. In some embodiments, N4 is G. In some embodiments, N4 is U. In some embodiments, N5 is A. In some embodiments, N5 is C. In some embodiments, N5 is G. In some embodiments, N5 is U. It will be understood that, each of the embodiments described above and herein (e.g., for N1 through N5) may be taken singly or in combination and / or may be combined with other embodiments of variables described above and herein (e.g., 5' caps).

[0321] In some embodiments, a cap proximal sequence comprises A1 and G2 of the Cap1 structure, and a sequence comprising: A3A4U5 at positions +3, +4 and +5 respectively of the nucleotide sequence. 3. 5’ UTR

[0322] In some embodiments, an RNA utilized in accordance with the present disclosure comprises a 5'-UTR. In some embodiments, a 5’-UTR may comprise a plurality of distinct sequence elements; in some embodiments, such plurality may be or comprise multiple copies of one or more particular sequence elements (e.g., as may be from a particular source or otherwise known as a functional or characteristic sequence element). In some embodiments, a 5’ UTR comprises multiple different sequence elements.

[0323] The term “untranslated region” or “UTR” is commonly used in the art to refer to a region in a DNA molecule which is transcribed but is not translated into an amino acid sequence, or to the corresponding region in an RNA polynucleotide. An untranslated region (UTR) can be present 5' (upstream) of an open reading frame (5'-UTR) and / or 3' (downstream) of an open reading frame (3'-UTR). As used herein, the terms “five prime untranslated region” or “5' UTR” refer to a sequence of a nucleotide sequence between the 5' end of the nucleotide sequence (e.g., a transcription start site) and a start codon of a coding region of the nucleotide sequence. In somePage 131 of 182 12197519v1embodiments, “5' UTR” refers to a sequence of a nucleotide sequence that begins at the 5' end of the nucleotide sequence (e.g., a transcription start site) and ends one nucleotide (nt) before a start codon (usually AUG) of a coding region of the nucleotide sequence, e.g., in its natural context. In some embodiments, a 5' UTR comprises a Kozak sequence. A 5'-UTR is downstream of the 5'-cap (if present), e.g., directly adjacent to the 5'-cap. In some embodiments, a 5' UTR disclosed herein comprises a cap proximal sequence, e.g., as defined and described herein. In some embodiments, a cap proximal sequence comprises a sequence adjacent to a 5' cap.

[0324] Exemplary 5' UTRs include a human alpha globin (hAg) 5'UTR or a fragment thereof, a TEV 5' UTR or a fragment thereof, a HSP705' UTR or a fragment thereof, or a c-Jun 5' UTR or a fragment thereof.

[0325] In some embodiments, an RNA disclosed herein comprises a hAg 5' UTR or a fragment thereof.

[0326] In some embodiments, an RNA disclosed herein comprises a 5' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 5' UTR with the sequence GGAATAAAAGTCTCAACACAACATATACAAAACAAACGAATCTCAAGCAATCAAGCATTCTACTTCTATTGCAGCAATTTAAATCA TTTCTTTTAAAGCAAAAGCAATTTTCTGAAAATTTTCACCATTTACGAACGATAGC (SEQ ID NO: 13). In some embodiments, an RNA disclosed herein comprises a 5' UTR having the sequence set forth in SEQ ID NO: 13.

[0327] In some embodiments, an RNA disclosed herein comprises a 5' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 5' UTR with the sequence AGAATAAACTAGTATTCTTCTGGTCCCCACAGACTCAGAGAGAACCCGCCACC (SEQ ID NO: 194). In some embodiments, an RNA disclosed herein comprises a 5' UTR having the sequence set forth in SEQ ID NO: 194.

[0328] In some embodiments, an RNA disclosed herein comprises a 5' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 5’ UTR with the sequence AACUAGUAUUCUUCUGGUCCCCACAGACUCAGAGAGAACCCGCCACC (SEQ ID NO: 195), corresponding to hAg-Kozak 5'UTR. In some embodiments, an RNA disclosed herein comprises a 5' UTR having the sequence set forth in SEQ ID NO: 195). 4. PolyA Tail

[0329] In some embodiments, a polynucleotide (e.g., DNA, RNA) disclosed herein comprises a polyadenylate (polyA) sequence, e.g., as described herein. In some embodiments, a polyA sequence is situated downstream of a 3'- UTR, e.g., adjacent to a 3'-UTR.

[0330] As used herein, the term “poly(A) sequence” or “poly-A tail” refers to an uninterrupted or interrupted sequence of adenylate residues which is typically located at the 3'-end of an RNA polynucleotide. Poly(A) sequences are known to those of skill in the art and may follow the 3’-UTR in the RNAs described herein. An uninterrupted poly(A) sequence is characterized by consecutive adenylate residues. In nature, an uninterrupted poly(A) sequence is typical. In some embodiments, polynucleotides disclosed herein comprise an uninterrupted poly(A) sequence. In some embodiments, polynucleotides disclosed herein comprise interrupted poly(A) sequence. In some embodiments, RNAs disclosed herein can have a poly(A) sequence attached to the free 3'-end of the RNA by a template- independent RNA polymerase after transcription or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase.Page 132 of 182 12197519v1

[0331] It has been demonstrated that a poly(A) sequence of about 120 A nucleotides has a beneficial influence on the levels of RNA in transfected eukaryotic cells, as well as on the levels of protein that are translated from an open reading frame that is present upstream (5') of the poly(A) sequence (Holtkamp et al., 2006, Blood, vol. 108, pp. 4009-4017, which is herein incorporated by reference).

[0332] In some embodiments, a poly(A) sequence in accordance with the present disclosure is not limited to a particular length; in some embodiments, a poly(A) sequence is any length. In some embodiments, a poly(A) sequence comprises, essentially consists of, or consists of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 A nucleotides, and, in particular, about 120 A nucleotides. In this context, "essentially consists of" means that most nucleotides in the poly(A) sequence, typically at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by number of nucleotides in the poly(A) sequence are A nucleotides, but permits that remaining nucleotides are nucleotides other than A nucleotides, such as U nucleotides (uridylate), G nucleotides (guanylate), or C nucleotides (cytidylate). In this context, "consists of" means that all nucleotides in the poly(A) sequence, i.e., 100% by number of nucleotides in the poly(A) sequence, are A nucleotides. The term “A nucleotide” or “A” refers to adenylate.

[0333] In some embodiments, a poly(A) sequence is attached during RNA transcription, e.g., during preparation of in vitro transcribed RNA, based on a DNA template comprising repeated dT nucleotides (deoxythymidylate) in the strand complementary to the coding strand. The DNA sequence encoding a poly(A) sequence (coding strand) is referred to as a poly(A) cassette.

[0334] In some embodiments, the poly(A) cassette present in the coding strand of DNA essentially consists of dA nucleotides, but is interrupted by a random sequence of the four nucleotides (dA, dC, dG, and dT). Such a random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length. Such a cassette is disclosed in WO 2016 / 005324 A1, hereby incorporated by reference. Any poly(A) cassette disclosed in WO 2016 / 005324 A1, which is incorporated herein by reference in its entirety, may be used in accordance with the present disclosure. A poly(A) cassette that essentially consists of dA nucleotides, but is interrupted by a random sequence having an equal distribution of the four nucleotides (dA, dC, dG, dT) and having a length of e.g., 5 to 50 nucleotides shows, at the DNA level, constant propagation of plasmid DNA in E. coli and is still associated, at the RNA level, with the beneficial properties with respect to supporting RNA stability and translational efficiency is encompassed. In some embodiments, the poly(A) sequence contained in an RNA polynucleotide described herein essentially consists of A nucleotides, but is interrupted by a random sequence of the four nucleotides (A, C, G, U). Such a random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length.

[0335] In some embodiments, no nucleotides other than A nucleotides flank a poly(A) sequence at its 3'-end, i.e., the poly(A) sequence is not masked or followed at its 3'-end by a nucleotide other than A.

[0336] In some embodiments, the poly(A) sequence may comprise at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence may essentially consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence may consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300,Page 133 of 182 12197519v1up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence comprises at least 100 nucleotides. In some embodiments, the poly(A) sequence comprises about 150 nucleotides. In some embodiments, the poly(A) sequence comprises about 120 nucleotides.

[0337] In some embodiments, a poly(A) sequence comprises a specific number of adenosines, such as about 50 or more, about 60 or more, about 70 or more, about 80 or more, about 90 or more, about 100 or more, about 120, (A)AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAA (SEQ ID NO: 16), or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 16.

[0339] In some embodiments, RNA comprises a poly(A) sequence comprising the nucleotide sequence of: AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGCAUAUGACUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 196), or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 196. In some embodiments, a poly(A) sequence comprises a plurality of A residues interrupted by a linker. In some embodiments, a linker comprises the nucleotide sequence GCAUAUGAC. 5. 3' UTR

[0340] In some embodiments, an RNA utilized in accordance with the present disclosure comprises a 3'-UTR. As used herein, the terms “three prime untranslated region,” “3' untranslated region,” or “3' UTR” refer to a sequence of an RNA molecule that begins following a stop codon of a coding region of an open reading frame sequence. In some embodiments, the 3' UTR begins immediately after a stop codon of a coding region of an open reading frame sequence, e.g., in its natural context. In other embodiments, the 3' UTR does not begin immediately after stop codon of the coding region of an open reading frame sequence, e.g., in its natural context. In some embodiments, the term “3'-UTR” preferably does not include the poly(A) sequence. Thus, the 3'-UTR is upstream of the poly(A) sequence (if present), e.g., directly adjacent to the poly(A) sequence.

[0341] In some embodiments, an RNA disclosed herein comprises a 3'TR comprising an F element and / or an I element. In some embodiments, a 3' UTR or a proximal sequence thereto comprises a restriction site. In some embodiments, a restriction site is a BamHI site. In some embodiments, a restriction site is an XhoI site.

[0342] In some embodiments, an RNA construct comprises an F element. In some embodiments, an F element sequence is a 3' UTR of amino-terminal enhancer of split (AES).

[0343] In some embodiments, an RNA disclosed herein comprises a 3' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 3’ UTR with the sequence ofPage 134 of 182 12197519v1CTGGTACTGCATGCACGCAATGCTAGCTGCCCCTTTCCCGTCCTGGGTACCCCGAGTCTCCCCCGACCTCGGGTCCCAGGTATG CTCCCACCTCCACCTGCCCCACTCACCACCTCTGCTAGTTCCAGACACCTCCCAAGCACGCAGCAATGCAGCTCAAAACGCTTAG CCTAGCCACACCCCCACGGGAAACAGCAGTGATTAACCTTTAGCAATAAACGAAAGTTTAACTAAGCTATACTAACCCCAGGGTT GGTCAATTTCGTGCCAGCCACACC (SEQ ID NO: 186). In some embodiments, an RNA disclosed herein comprises a 3' UTR with the sequence of SEQ ID NO: 186.

[0344] In some embodiments, an RNA disclosed herein comprises a 3' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 3’ UTR with the sequence of CTAGTAGTGACTGACTAGGATCTGGTTACCACTAAACCAGCCTCAAGAACACCCGAATGGAGTCTCTAAGCTACATAATACCAAC TTACACTTACAAAATGTTGTCCCCCAAAATGTAGCCATTCGTATCTGCTCCTAATAAAAAGAAAGTTTCTTCACATTCT (SEQ ID NO: 198). In some embodiments, an RNA disclosed herein comprises a 3' UTR with the sequence set forth in SEQ ID NO: 198.

[0345] In some embodiments, a 3' UTR is an FI element as described in WO2017 / 060314, which is herein incorporated by reference in its entirety.

[0346] In some embodiments, a polyribonucleotide as described herein comprises a 5′ untranslated region, 3′ untranslated region, poly-A tail, 5’ cap, or a combination thereof.

[0347] In some embodiments, the nucleotide sequences disclosed herein are optimized according to techniques generally known in the art. For example, codon optimization is an approach in gene engineering to improve gene expression by changing synonymous codons based on an organism's codon bias. As it is well-known in the art, nucleotide changes can be made throughout a nucleotide sequence of interest based on an organism’s codon usage bias to increase translational efficiency and thus protein expression without altering the sequence of the protein. Nucleoside Modification

[0348] In some embodiments, the present disclosure provides compositions comprising modified RNAs and methods of use thereof. In some embodiments, the modified RNA comprises one or more modified nucleoside residues. For example, in some embodiments, an RNA comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 2, or Table 4, comprises one or more modified nucleoside residues.

[0349] In some embodiments, an RNA as described herein refers to a messenger RNA.

[0350] In some embodiments, all uridine residues are modified as described herein. In some embodiments, one or more of the RNAs as described herein are nucleoside modified RNAs. In other embodiments, two or more of the RNAs as described herein are nucleoside modified RNAs. In other embodiments, three or more of the RNAs as described herein are nucleoside modified RNAs.

[0351] In some embodiments, the modified nucleoside of the methods and compositions of the present disclosure is m5C (5-methylcytidine). In another embodiment, the modified nucleoside is m5U (5-methyluridine). In another embodiment, the modified nucleoside is m6A (N6-methyladenosine). In another embodiment, the modifiedPage 135 of 182 12197519v1nucleoside is s2U (2-thiouridine). In another embodiment, the modified nucleoside is Ψ (pseudouridine). In another embodiment, the modified nucleoside is Um (2'-O-methyluridine).

[0352] In some embodiments, a modified nucleoside is m1A (1-methyladenosine), m2A (2-methyladenosine), m6A (N6-methyladenosine), Am (2'-O-methyladenosine), ms2m6A (2-methylthio-N6-methyladenosine), i6A (N6- isopentenyladenosine), ms2i6A (2-methylthio-N6-isopentenyladenosine), io6A (N6-(cis-hydroxyisopentenyl)adenosine), ms2io6A (2-methylthio-N6-(cis-hydroxyisopentenyl) adenosine), g6A (N6-glycinylcarbamoyladenosine), t6A (N6- threonylcarbamoyladenosine), ms2t6A (2-methylthio-N6-threonyl carbamoyladenosine), m6t6A (N6-methyl-N6- threonylcarbamoyladenosine), hn6A (N6-hydroxynorvalylcarbamoyladenosine), ms2hn6A (2-methylthio-N6- hydroxynorvalyl carbamoyladenosine), Ar(p) (2'-O-ribosyladenosine (phosphate)), I (inosine), m1I (1-methylinosine), m1Im (1,2'-O-dimethylinosine), m3C (3-methylcytidine), m5C (5-methylcytidine), Cm (2'-O-methylcytidine), s2C (2- thiocytidine), ac4C (N4-acetylcytidine), f5C (5-formylcytidine), m5Cm (5,2'-O-dimethylcytidine), ac4Cm (N4-acetyl-2'- O-methylcytidine), k2C (lysidine), m1G (1-methylguanosine), m2G (N2-methylguanosine), m7G (7-methylguanosine), Gm (2'-O-methylguanosine), m22G (N2,N2-dimethylguanosine), m2Gm (N2,2'-O-dimethylguanosine), m22Gm (N2,N2,2'-O-trimethylguanosine), Gr(p) (2'-O-ribosylguanosine (phosphate)), yW (wybutosine), o2yW (peroxywybutosine), OHyW (hydroxywybutosine), OHyW* (undermodified hydroxywybutosine), imG (wyosine), mimG (methylwyosine), Q (queuosine), oQ (epoxyqueuosine), galQ (galactosyl-queuosine), manQ (mannosyl- queuosine), preQ0 (7-cyano-7-deazaguanosine), preQ1 (7-aminomethyl-7-deazaguanosine), G+(archaeosine), Ψ (pseudouridine), D (dihydrouridine), m5U (5-methyluridine), Um (2'-O-methyluridine), m5Um (5,2'-O-dimethyluridine), m1Ψ (1-methylpseudouridine), Ψm (2'-O-methylpseudouridine), s2U (2-thiouridine), s4U (4-thiouridine), m5s2U (5- methyl-2-thiouridine), s2Um (2-thio-2'-O-methyluridine), acp3U (3-(3-amino-3-carboxypropyl)uridine), ho5U (5- hydroxyuridine), mo5U (5-methoxyuridine), cmo5U (uridine 5-oxyacetic acid), mcmo5U (uridine 5-oxyacetic acid methyl ester), chm5U (5-(carboxyhydroxymethyl)uridine), mchm5U (5-(carboxyhydroxymethyl)uridine methyl ester), mcm5U (5-methoxycarbonylmethyluridine), mcm5Um (5-methoxycarbonylmethyl-2'-O-methyluridine), mcm5s2U (5- methoxycarbonylmethyl-2-thiouridine), nm5s2U (5-aminomethyl-2-thiouridine), mnm5U (5- methylaminomethyluridine), mnm5s2U (5-methylaminomethyl-2-thiouridine), mnm5se2U (5-methylaminomethyl-2- selenouridine), ncm5U (5-carbamoylmethyluridine), ncm5Um (5-carbamoylmethyl-2'-O-methyluridine), cmnm5U (5- carboxymethylaminomethyluridine), cmnm5Um (5-carboxymethylaminomethyl- 2'-O-methyluridine), cmnm5s2U (5- carboxymethylaminomethyl-2-thiouridine), m62A (N6,N6-dimethyladenosine), Im (2'-O-methylinosine), m4C (N4- methylcytidine), m4Cm (N4,2'-O-dimethylcytidine), hm5C (5-hydroxymethylcytidine), m3U (3-methyluridine), m1acp3Ψ (1-methyl-3-(3-amino-3-carboxypropyl) pseudouridine), cm5U (5-carboxymethyluridine), m6Am (N6,2'-O- dimethyladenosine), m62Am (N6,N6,2'-O-trimethyladenosine), m2,7G (N2,7-dimethylguanosine), m2,2,7G (N2,N2,7- trimethylguanosine), m3Um (3,2'-O-dimethyluridine), m5D (5-methyldihydrouridine), m3Ψ (3-methylpseudouridine), f5Cm (5-formyl-2'-O-methylcytidine), m1Gm (1,2'-O-dimethylguanosine), m1Am (1,2'-O-dimethyladenosine), τm5U (5- taurinomethyluridine), τm5s2U (5-taurinomethyl-2-thiouridine), imG-14 (4-demethylwyosine), imG2 (isowyosine), ac6A (N6-acetyladenosine), inm5U (5-(isopentenylaminomethyl)uridine), inm5s2U (5-(isopentenylaminomethyl)- 2- thiouridine), inm5Um (5-(isopentenylaminomethyl)- 2'-O-methyluridine), m2,7Gm (N2,7,2'-O-trimethylguanosine), m42Cm (N4,N4,2'-O-trimethylcytidine), C+(agmatidine), m8A (8-methyladenosine), gmnm5s2U (geranylated 5- methylaminomethyl-2-thiouridine), gcmnm5s2U (geranylated 5-carboxymethylaminomethyl-2-thiouridine), or cnm5U (5-cyanomethyl-uridine).Page 136 of 182 12197519v1

[0353] In some embodiments, modified nucleoside residues are pseudouridine or pseudouridine family residues.

[0354] In some embodiments, the modified RNA comprises pseudouridine residues. In some embodiments, pseudouridine refers to the C-glycoside isomer of the nucleoside uridine. In some embodiments, pseudouridine residues comprise m1acp3Ψ (1-methyl-3-(3-amino-5-carboxypropyl)pseudouridine, m1Ψ (1-methylpseudouridine), Ψm (2′-O-methylpseudouridine, m5D (5-methyldihydrouridine), m3Ψ (3-methylpseudouridine), or a combination thereof. In some embodiments, said pseudouridine residues comprise 1-methylpseudouridine residues instead of uridine.

[0355] In some embodiments, the modified nucleoside residues are pseudouridine analogues. In some embodiments, a "pseudouridine analog" is any modification, variant, isoform or derivative of pseudouridine. For example, pseudouridine analogs include but are not limited to 1-carboxymethyl-pseudouridine, 1-propynyl- pseudouridine, 1-taurinomethyl-pseudouridine, 1-taurinomethyl-4-thio-pseudouridine, 1-methylpseudouridine (m1Ψ), 1-methyl-4-thio-pseudouridine (m1s4Ψ), 4-thio-1-methyl-pseudouridine, 3-methyl-pseudouridine (m3Ψ), 2-thio-1- methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydropseudouridine, 2-thio-dihydropseudouridine, 2-methoxyuridine, 2-methoxy-4-thio-uridine, 4-methoxy- pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl-pseudouridine, 1-methyl-3-(3-amino-3- carboxypropyl)pseudouridine (acp3Ψ), and 2'-O-methyl-pseudouridine (Ψm).

[0356] In some embodiments, the modified nucleobase is a modified uracil. Exemplary nucleobases and nucleosides having a modified uracil include pseudouridine (Ψ), pyridin-4-one ribonucleoside, 5-aza-uridine, 6-aza- uridine, 2-thio-5-aza-uridine, 2-thio-uridine (s2U), 4-thio-uridine (s4U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5- hydroxy-uridine (ho5U), 5-aminoallyl-uridine, 5-halo-uridine (e.g., 5-iodo-uridine or 5-bromo-uridine), 3-methyl- uridine (m3U), 5-methoxy-uridine (mo5U), uridine 5-oxyacetic acid (cmo5U), uridine 5-oxyacetic acid methyl ester (mcmo5U), 5-carboxymethyl-uridine (cm5U), 1-carboxymethyl-pseudouridine, 5-carboxyhydroxymethyl-uridine (chm5U), 5-carboxyhydroxymethyl-uridine methyl ester (mchm5U), 5-methoxycarbonylmethyl-uridine (mcm5U), 5- methoxycarbonylmethyl-2-thio-uridine (mcm5s2U), 5-aminomethyl-2-thio-uridine (nm5s2U), 5-methylaminomethyl- uridine (mnm5U), 5-methylaminomethyl-2-thio-uridine (mnm5s2U), 5-methylaminomethyl-2-seleno-uridine (mnm5se2U), 5-carbamoylmethyl-uridine (ncm5U), 5-carboxymethylaminomethyl-uridine (cmnm5U), 5- carboxymethylaminomethyl-2-thio-uridine (cmnm5s2U), 5-propynyl-uridine, 1-propynyl-pseudouridine, 5- taurinomethyl-uridine (τcm5U), 1-taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uridine (τrm5s2U), 1- taurinomethyl-4-thio-pseudouridine, 5-methyl-uridine (m5U, i.e., having the nucleobase deoxythymine), 1- methylpseudouridine (m1Ψ), 5-methyl-2-thio-uridine (m5s2U), 1-methyl-4-thio-pseudouridine (m1s4Ψ), 4-thio-1- methyl-pseudouridine, 3-methyl-pseudouridine (m3Ψ), 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza- pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine (D), dihydropseudouridine, 5,6-dihydrouridine, 5-methyl-dihydrouridine (m5D), 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2-methoxy-uridine, 2-methoxy-4- thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl-pseudouridine (also known as 1- methylpseudouridine (m1Ψ), 3-(3-amino-3-carboxypropyl)uridine (acp3U), 1-methyl-3-(3-amino-3- carboxypropyl)pseudouridine (acp3Ψ), 5-(isopentenylaminomethyl)uridine (inm5U), 5-(isopentenylaminomethyl)-2- thio-uridine (inm5s2U), α-thio-uridine, 2'-O-methyl-uridine (Um), 5,2'-O-dimethyl-uridine (m5Um), 2'-O-methyl- pseudouridine (Ψm), 2-thio-2'-O-methyl-uridine (s2Um), 5-methoxycarbonylmethyl-2'-O-methyl-uridine (mcm5Um), 5-Page 137 of 182 12197519v1carbamoylmethyl-2'-β-methyl-uridine (ncm5Um), 5-carboxymethylaminomethyl-2'-O-methyl-uridine (cmnm5Um), 3,2'- O-dimethyl-uridine (m3Um), 5-(isopentenylaminomethyl)-2'-β-methyl-uridine (inm5Um), 1-thio-uridine, deoxythymidine, 2'-F-ara-uridine, 2'-F-uridine, 2'-OH-ara-uridine, 5-(2-carbomethoxyvinyl)uridine, and 5-[3-(1-E- propenylamino)uridine.

[0357] In some embodiments, the modified nucleobase is a modified cytosine. Exemplary nucleobases and nucleosides having a modified cytosine include 5-aza-cytidine, 6-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine (m3C), N4-acetyl-cytidine (ac4C), 5-formyl-cytidine (f5C), N4-methyl-cytidine (m4C), 5-methyl-cytidine (m5C), 5-halo- cytidine (e.g., 5-iodo-cytidine), 5-hydroxymethyl-cytidine (hm5C), 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo-pseudoisocytidine, 2-thio-cytidine (s2C), 2-thio-5-methyl-cytidine, 4-thio-pseudoisocytidine, 4-thio-1-methyl- pseudoisocytidine, 4-thio-1-methyl-1-deaza-pseudoisocytidine, 1-methyl-1-deaza-pseudoisocytidine, zebularine, 5- aza-zebularine, 5-methyl-zebularine, 5-aza-2-thio-zebularine, 2-thio-zebularine, 2-methoxy-cytidine, 2-methoxy-5- methyl-cytidine, 4-methoxy-pseudoisocytidine, 4-methoxy-1-methyl-pseudoisocytidine, lysidine (k2C), α-thio-cytidine, 2'-O-methyl-cytidine (Cm), 5,2'-O-dimethyl-cytidine (m5Cm), N4-acetyl-2'-O-methyl-cytidine (ac4Cm), N4,2'-O- dimethyl-cytidine (m4Cm), 5-formyl-2'-O-methyl-cytidine (f5Cm), N4,N4,2'-O-trimethyl-cytidine (m42Cm), 1-thio- cytidine, 2'-F-ara-cytidine, 2'-F-cytidine, and 2'-OH-ara-cytidine.

[0358] In some embodiments, the modified nucleobase is a modified adenine. Exemplary nucleobases and nucleosides having a modified adenine include 2-amino-purine, 2,6-diaminopurine, 2-amino-6-halo-purine (e.g., 2- amino-6-chloro-purine), 6-halo-purine (e.g., 6-chloro-purine), 2-amino-6-methyl-purine, 8-azido-adenosine, 7-deaza- adenine, 7-deaza-8-aza-adenine, 7-deaza-2-amino-purine, 7-deaza-8-aza-2-amino-purine, 7-deaza-2,6- diaminopurine, 7-deaza-8-aza-2,6-diaminopurine, 1-methyl-adenosine (m1A), 2-methyl-adenine (m2A), N6-methyl- adenosine (m6A), 2-methylthio-N6-methyl-adenosine (ms2m6A), N6-isopentenyl-adenosine (i6A), 2-methylthio-N6- isopentenyl-adenosine (ms2i6A), N6-(cis-hydroxyisopentenyl)adenosine (io6A), 2-methylthio-N6-(cis- hydroxyisopentenyl)adenosine (ms2io6A), N6-glycinylcarbamoyl-adenosine (g6A), N6-threonylcarbamoyl-adenosine (t6A), N6-methyl-N6-threonylcarbamoyl-adenosine (m6t6A), 2-methylthio-N6-threonylcarbamoyl-adenosine (ms2g6A), N6,N6-dimethyl-adenosine (m62A), N6-hydroxynorvalylcarbamoyl-adenosine (hn6A), 2-methylthio-N6- hydroxynorvalylcarbamoyl-adenosine (ms2hn6A), N6-acetyl-adenosine (ac6A), 7-methyl-adenine, 2-methylthio- adenine, 2-methoxy-adenine, α-thio-adenosine, 2'-O-methyl-adenosine (Am), N6,2'-O-dimethyl-adenosine (m6Am), N6,N6,2'-O-trimethyl-adenosine (m62Am), 1,2'-O-dimethyl-adenosine (m1Am), 2'-β-ribosyladenosine (phosphate) (Ar(p)), 2-amino-N6-methyl-purine, 1-thio-adenosine, 8-azido-adenosine, 2'-F-ara-adenosine, 2'-F-adenosine, 2'-OH- ara-adenosine, and N6-(19-amino-pentaoxanonadecyl)-adenosine.

[0359] In some embodiments, the modified nucleobase is a modified guanine. Exemplary nucleobases and nucleosides having a modified guanine include inosine (I), 1-methyl-inosine (m1I), wyosine (imG), methylwyosine (mimG), 4-demethyl-wyosine (imG-14), isowyosine (imG2), wybutosine (yW), peroxywybutosine (o2yW), hydroxywybutosine (OHyW), undermodified hydroxywybutosine (OHyW*), 7-deaza-guanosine, queuosine (Q), epoxyqueuosine (oQ), galactosyl-queuosine (galQ), mannosyl-queuosine (manQ), 7-cyano-7-deaza-guanosine (preQ0), 7-aminomethyl-7-deaza-guanosine (preQ1), archaeosine (G+), 7-deaza-8-aza-guanosine, 6-thio-guanosine, 6-thio-7-deaza-guanosine, 6-thio-7-deaza-8-aza-guanosine, 7-methyl-guanosine (m7G), 6-thio-7-methyl-guanosine, 7-methyl-inosine, 6-methoxy-guanosine, 1-methyl-guanosine (m1G), N2-methyl-guanosine (m2G), N2,N2-dimethyl-Page 138 of 182 12197519v1guanosine (m22G), N2,7-dimethyl-guanosine (m2,7G), N2,N2,7-dimethyl-guanosine (m2,2,7G), 8-oxo-guanosine, 7- methyl-8-oxo-guanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, N2,N2-dimethyl-6-thio-guanosine, α-thio-guanosine, 2'-O-methyl-guanosine (Gm), N2-methyl-2'-O-methyl-guanosine (m2Gm), N2,N2-dimethyl-2'-O- methyl-guanosine (m22Gm), 1-methyl-2'-O-methyl-guanosine (m1Gm), N2,7-dimethyl-2'-O-methyl-guanosine (m2'7Gm), 2'-O-methyl-inosine (Im), 1,2'-O-dimethyl-inosine (m1Im), and 2'-O-ribosylguanosine (phosphate) (Gr(p)).

[0360] The nucleobase of the nucleotide can be independently selected from a purine, a pyrimidine, a purine or pyrimidine analog. For example, the nucleobase can each be independently selected from adenine, cytosine, guanine, uracil, or hypoxanthine. In another embodiment, the nucleobase can also include, for example, naturally-occurring and synthetic derivatives of a base, including pyrazolo[3,4-d]pyrimidines, 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-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo (e.g., 8-bromo), 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, 8-azaguanine and 8-azaadenine, deazaguanine, 7-deazaguanine, 3-deazaguanine, deazaadenine, 7- deazaadenine, 3-deazaadenine, pyrazolo[3,4-d]pyrimidine, imidazo[1,5-a]1,3,5 triazinones, 9-deazapurines, imidazo[4,5-d]pyrazines, thiazolo[4,5-d]pyrimidines, pyrazin-2-ones, 1,2,4-triazine, pyridazine; and 1,3,5 triazine. When the nucleotides are depicted using the shorthand A, G, C, T or U, each letter refers to the representative base and / or derivatives thereof, e.g., A includes adenine or adenine analogs, e.g., 7-deaza adenine). Modifications on the Internucleoside Linkage

[0361] The modified nucleotides, which may be incorporated into a polynucleotide, primary construct, or RNA molecule, can be modified on the internucleoside linkage (e.g., phosphate backbone). Herein, in the context of the polynucleotide backbone, the phrases "phosphate" and "phosphodiester" are used interchangeably. Backbone phosphate groups can be modified by replacing one or more of the oxygen atoms with a different substituent. Further, the modified nucleosides and nucleotides can include the wholesale replacement of an unmodified phosphate moiety with another internucleoside linkage as described herein. Examples of modified phosphate groups include, but are not limited to, phosphorothioate, phosphoroselenates, boranophosphates, boranophosphate esters, hydrogen phosphonates, phosphoramidates, phosphorodiamidates, alkyl or aryl phosphonates, and phosphotriesters. Phosphorodithioates have both non-linking oxygens replaced by sulfur. The phosphate linker can also be modified by the replacement of a linking oxygen with nitrogen (bridged phosphoramidates), sulfur (bridged phosphorothioates), and carbon (bridged methylene-phosphonates).

[0362] The α-thio substituted phosphate moiety is provided to confer stability to RNA and DNA polymers through the unnatural phosphorothioate backbone linkages. Phosphorothioate DNA and RNA have increased nuclease resistance and subsequently a longer half-life in a cellular environment. Phosphorothioate linked polynucleotides, primary constructs, or modified RNA molecules are expected to also reduce the innate immune response through weaker binding / activation of cellular innate immune molecules.Page 139 of 182 12197519v1

[0363] In specific embodiments, a modified nucleoside includes an alpha-thio-nucleoside (e.g., 5'-O-(1- thiophosphate)-adenosine, 5'-O-(1-thiophosphate)-cytidine (α-thio-cytidine), 5'-O-(1-thiophosphate)-guanosine, 5'-O- (1-thiophosphate)-uridine, or 5'-O-(1-thiophosphate)-pseudouridine).

[0364] Other internucleoside linkages that may be employed according to the present disclosure, including internucleoside linkages which do not contain a phosphorous atom, are described herein below. Combinations of Modified Sugars, Nucleobases, and Internucleoside Linkages

[0365] The polynucleotides, primary constructs, and modified RNA of the disclosure can include a combination of modifications to the sugar, the nucleobase, and / or the internucleoside linkage.

[0366] In another embodiment, the purified preparation of RNA, oligoribonucleotide, or polyribonucleotide of the methods and compositions of the present disclosure comprises a combination of two or more of the above-described modifications. In another embodiment, the purified preparation of the RNA or oligoribonucleotide comprises a combination of three or more of the above-described modifications. In another embodiment, the purified preparation of the RNA or oligoribonucleotide comprises a combination of more than three of the above-described modifications.

[0367] In some embodiments, the modified RNAs comprise in vitro-synthesized modified RNAs.

[0368] In some embodiments, the present disclosure comprises one or more modified RNAs encoding an HSV glycoprotein. In some embodiments, the modified RNA comprises pseudouridine or pseudouridine family residues. In another embodiment, the modified RNAs of the present disclosure are capable of directing protein expression of HSV glycoproteins encoded thereon.

[0369] In another embodiment, the present disclosure provides an in vitro-transcribed RNA molecule encoding an HSV glycoprotein, comprising a pseudouridine. In another embodiment, the present disclosure provides a synthetic RNA molecule encoding an HSV glycoprotein, comprising a pseudouridine.

[0370] In another embodiment, an in vitro-transcribed RNA molecule of the methods and compositions of the present disclosure is synthesized by T7 phage RNA polymerase. In another embodiment, the molecule is synthesized by SP6 phage RNA polymerase. In another embodiment, the molecule is synthesized by T3 phage RNA polymerase. In another embodiment, the molecule is synthesized by a polymerase selected from the above polymerases. In another embodiment, the RNA is synthesized chemically on a column similar to DNA.

[0371] In another embodiment, the nucleoside that is modified in an RNA, oligoribonucleotide, or polyribonucleotide of the methods and compositions of the present disclosure is uridine (U). In another embodiment, the modified nucleoside is cytidine (C). In another embodiment, the modified nucleoside is adenine (A). In another embodiment the modified nucleoside is guanine (G).

[0372] In another embodiment, the RNA of the methods and compositions of the present disclosure further comprises a poly-A tail. In another embodiment, the RNA of the methods and compositions of the present disclosure does not comprise a poly-A tail. Each possibility represents a separate embodiment of the present disclosure.

[0373] In another embodiment, the RNA of the methods and compositions of the present disclosure comprises an m7GpppG cap. In another embodiment, the RNA of the methods and compositions of the present disclosure does not comprise an m7GpppG cap. In another embodiment, the RNA of the methods and compositions of the presentPage 140 of 182 12197519v1disclosure comprises a 3′-O-methyl-m7GpppG. In another embodiment, the RNA of methods and composition of the present disclosure comprise a non-reversible cap analog, which, in some embodiments, is added during transcription of the RNA. In another embodiment, the RNA of methods and composition of the present disclosure comprise an anti-reverse cap analog. Each possibility represents a separate embodiment of the present disclosure.

[0374] In another embodiment, the RNA of the methods and compositions of the present disclosure further comprises a cap-independent translational enhancer. In another embodiment, the RNA of the methods and compositions of the present disclosure does not comprise a cap-independent translational enhancer. In another embodiment, the cap-independent translational enhancer is a tobacco etch virus (TEV) cap-independent translational enhancer. In another embodiment, the cap-independent translational enhancer is any other cap-independent translational enhancer known in the art. Each possibility represents a separate embodiment of the present disclosure.

[0375] In some embodiments, “pseudouridine” refers to m1acp3Ψ (1-methyl-3-(3-amino-5- carboxypropyl)pseudouridine. In another embodiment, the term refers to m1Ψ (1-methylpseudouridine). In another embodiment, the term refers to Ψm (2′-O-methylpseudouridine. In another embodiment, the term refers to m5D (5- methyldihydrouridine). In another embodiment, the term refers to m3Ψ (3-methylpseudouridine). In another embodiment, the modified nucleoside is 4' (pseudouridine). In another embodiment, the term refers to a pseudouridine moiety that is not further modified. In another embodiment, the term refers to a monophosphate, diphosphate, or triphosphate of any of the above pseudouridines. In another embodiment, the term refers to any other pseudouridine known in the art. Each possibility represents a separate embodiment of the present disclosure.

[0376] In another embodiment, the modified RNA comprises a modified nucleoside, which in some embodiments, comprises m5C, m5U, m6A, s2U, Ψ, 2'-O-methyl-U, 2’-O-methylpseudouridine, or a combination thereof.

[0377] In another embodiment, the present disclosure provides a method for delivering a recombinant protein to a subject, the method comprising the step of contacting the subject with an RNA of the methods and compositions of the present disclosure, thereby delivering a recombinant protein to a subject.

[0378] In another embodiment, a method of the present disclosure comprises increasing the number, percentage, or frequency of modified uridine nucleosides in the RNA molecule to decrease immunogenicity or increase efficiency of translation. In some embodiments, the number of modified uridine residues in an RNA, oligoribonucleotide, or polyribonucleotide molecule determines the magnitude of the effects observed in the present disclosure.

[0379] In another embodiment, between 0.1% and 100% of the uridine residues in the modified RNAs of the methods and compositions of the present disclosure are modified (e.g. by the presence of pseudouridine). In another embodiment, 0.1% of the residues are modified. In another embodiment, 0.2%. In another embodiment, the fraction is 0.3%. In another embodiment, the fraction is 0.4%. In another embodiment, the fraction is 0.5%. In another embodiment, the fraction is 0.6%. In another embodiment, the fraction is 0.8%. In another embodiment, the fraction is 1%. In another embodiment, the fraction is 1.5%. In another embodiment, the fraction is 2%. In another embodiment, the fraction is 2.5%. In another embodiment, the fraction is 3%. In another embodiment, the fraction is 4%. In another embodiment, the fraction is 5%. In another embodiment, the fraction is 6%. In another embodiment, the fraction is 8%. In another embodiment, the fraction is 10%. In another embodiment, the fraction isPage 141 of 182 12197519v112%. In another embodiment, the fraction is 14%. In another embodiment, the fraction is 16%. In another embodiment, the fraction is 18%. In another embodiment, the fraction is 20%. In another embodiment, the fraction is 25%. In another embodiment, the fraction is 30%. In another embodiment, the fraction is 35%. In another embodiment, the fraction is 40%. In another embodiment, the fraction is 45%. In another embodiment, the fraction is 50%. In another embodiment, the fraction is 60%. In another embodiment, the fraction is 70%. In another embodiment, the fraction is 80%. In another embodiment, the fraction is 90%. In another embodiment, the fraction is 100%.

[0380] In another embodiment, the fraction is less than 5%. In another embodiment, the fraction is less than 3%. In another embodiment, the fraction is less than 1%. In another embodiment, the fraction is less than 2%. In another embodiment, the fraction is less than 4%. In another embodiment, the fraction is less than 6%. In another embodiment, the fraction is less than 8%. In another embodiment, the fraction is less than 10%. In another embodiment, the fraction is less than 12%. In another embodiment, the fraction is less than 15%. In another embodiment, the fraction is less than 20%. In another embodiment, the fraction is less than 30%. In another embodiment, the fraction is less than 40%. In another embodiment, the fraction is less than 50%. In another embodiment, the fraction is less than 60%. In another embodiment, the fraction is less than 70%

[0381] In another embodiment, 0.1% of the residues of a given uridine nucleotide are modified. In another embodiment, the fraction of the nucleotide is 0.2%. In another embodiment, the fraction is 0.3%. In another embodiment, the fraction is 0.4%. In another embodiment, the fraction is 0.5%. In another embodiment, the fraction is 0.6%. In another embodiment, the fraction is 0.8%. In another embodiment, the fraction is 1%. In another embodiment, the fraction is 1.5%. In another embodiment, the fraction is 2%. In another embodiment, the fraction is 2.5%. In another embodiment, the fraction is 3%. In another embodiment, the fraction is 4%. In another embodiment, the fraction is 5%. In another embodiment, the fraction is 6%. In another embodiment, the fraction is 8%. In another embodiment, the fraction is 10%. In another embodiment, the fraction is 12%. In another embodiment, the fraction is 14%. In another embodiment, the fraction is 16%. In another embodiment, the fraction is 18%. In another embodiment, the fraction is 20%. In another embodiment, the fraction is 25%. In another embodiment, the fraction is 30%. In another embodiment, the fraction is 35%. In another embodiment, the fraction is 40%. In another embodiment, the fraction is 45%. In another embodiment, the fraction is 50%. In another embodiment, the fraction is 60%. In another embodiment, the fraction is 70%. In another embodiment, the fraction is 80%. In another embodiment, the fraction is 90%. In another embodiment, the fraction is 100%.

[0382] In another embodiment, the fraction of the given uridine nucleotide is less than 8%. In another embodiment, the fraction is less than 10%. In another embodiment, the fraction is less than 5%. In another embodiment, the fraction is less than 3%. In another embodiment, the fraction is less than 1%. In another embodiment, the fraction is less than 2%. In another embodiment, the fraction is less than 4%. In another embodiment, the fraction is less than 6%. In another embodiment, the fraction is less than 12%. In another embodiment, the fraction is less than 15%. In another embodiment, the fraction is less than 20%. In another embodiment, the fraction is less than 30%. In another embodiment, the fraction is less than 40%. In another embodiment, the fraction is less than 50%. In another embodiment, the fraction is less than 60%. In another embodiment, the fraction is less than 70%.Page 142 of 182 12197519v1

[0383] In another embodiment, the terms “ribonucleotide,” “oligoribonucleotide,” and polyribonucleotide refers to, in some embodiments, compounds comprising nucleotides in which the sugar moiety is ribose. In another embodiment, the term includes both RNA and RNA derivates in which the backbone is modified. Numerous RNA backbone modifications are known in the art and contemplated in the present disclosure. In some embodiments, modified RNA is a PNA (peptide nucleic acid). PNA contain peptide backbones and nucleotide bases and are able to bind, in another embodiment, to both DNA and RNA molecules. In another embodiment, the nucleotide is modified by replacement of one or more phosphodiester bonds with a phosphorothioate bond. In another embodiment, the artificial nucleic acid contains any other variant of the phosphate backbone of native nucleic acids known in the art. Each nucleic acid derivative represents a separate embodiment of the present disclosure.

[0384] Methods for production of nucleic acids having modified backbones are well known in the art, and are described, for example in U.S. Pat. Nos. 5,723,335 and 5,663,153 issued to Hutcherson et al. and related PCT publication WO95 / 26204. Each method represents a separate embodiment of the present disclosure.

[0385] The nucleic acid of interest can be purified by any method known in the art, or any method to be developed, so long as the method of purification removes contaminants from the nucleic acid preparation and thereby substantially reduces the immunogenicity potential of the nucleic acid preparation. In some embodiments, the nucleic acid of interest is purified using high-performance liquid chromatography (HPLC). In another embodiment, the nucleic acid of interest is purified by contacting the nucleic acid of interest with the bacterial enzyme RNase III. In other various embodiments, any method of nucleic acid purification that substantially reduces the immunogenicity of the nucleic acid preparation can be used. Non-limiting examples of purification methods that can be used with the compositions and methods of the disclosure include liquid chromatography separation and enzyme digestion, each used alone or in any combination, simultaneously or in any order. Non-limiting examples of liquid chromatography separation include HPLC and fast protein liquid chromatography (FPLC). Materials useful in the HPLC and FPLC methods of the disclosure include, but are not limited to, cross-linked polystyrene / divinylbenzene (PS / DVB), PS / DVB-C18, PS / DVB-alkylated, Helix DNA columns (Varian), Eclipse dsDNA Analysis Columns (Agilent Technologies), Reverse-phase 5 (RPC-5) exchange material, DNAPac, ProSwift, and bio-inert UltiMate.RTM. 3000 Titanium columns (Dionex). Enzymes useful in the enzyme digestion methods of the disclosure include any enzyme able to digest any contaminant in a nucleic acid preparation of the disclosure, such as, for example a dsRNA contaminant, and include but are not limited to, RNase III, RNase V1, Dicer, and Chipper (see Fruscoloni et al., 2002, PNAS 100:1639) Non-limiting examples of assays for assessing the purity of the nucleic acid of interest include a dot- blot assay, a Northern blot assay, and a dendritic cell activation assay, as described elsewhere herein.

[0386] In another embodiment, the modified RNA of the methods and compositions of the present disclosure is significantly less immunogenic than an unmodified in vitro-synthesized RNA molecule with the same sequence. In another embodiment, the modified RNA molecule is 2-fold less immunogenic than its unmodified counterpart. In another embodiment, immunogenicity is reduced by a 3-fold factor. In another embodiment, immunogenicity is reduced by a 5-fold factor. In another embodiment, immunogenicity is reduced by a 7-fold factor. In another embodiment, immunogenicity is reduced by a 10-fold factor. In another embodiment, immunogenicity is reduced by a 15-fold factor. In another embodiment, immunogenicity is reduced by a fold factor. In another embodiment, immunogenicity is reduced by a 50-fold factor. In another embodiment, immunogenicity is reduced by a 100-foldPage 143 of 182 12197519v1factor. In another embodiment, immunogenicity is reduced by a 200-fold factor. In another embodiment, immunogenicity is reduced by a 500-fold factor. In another embodiment, immunogenicity is reduced by a 1000-fold factor. In another embodiment, immunogenicity is reduced by a 2000-fold factor. In another embodiment, immunogenicity is reduced by another fold difference.

[0387] In another embodiment, “significantly less immunogenic” refers to a detectable decrease in immunogenicity. In another embodiment, the term refers to a fold decrease in immunogenicity (e.g., 1 of the fold decreases enumerated above). In another embodiment, the term refers to a decrease such that an effective amount of the modified RNA can be administered without triggering a detectable immune response. In another embodiment, the term refers to a decrease such that the modified RNA can be repeatedly administered without eliciting an immune response sufficient to detectably reduce expression of the recombinant protein. In another embodiment, the decrease is such that the modified RNA can be repeatedly administered without eliciting an immune response sufficient to eliminate detectable expression of the recombinant protein.

[0388] Methods of determining immunogenicity are well known in the art, and described in detail in U. S. Patent 8,278,036 which is hereby incorporated by reference herein.

[0389] In another embodiment, the modified RNA of the methods and compositions of the present disclosure is translated in the cell more efficiently than an unmodified RNA molecule with the same sequence. In another embodiment, the modified RNA exhibits enhanced ability to be translated by a target cell. In another embodiment, translation is enhanced by a factor of 2-fold relative to its unmodified counterpart. In another embodiment, translation is enhanced by a 3-fold factor. In another embodiment, translation is enhanced by a 5-fold factor. In another embodiment, translation is enhanced by a 7-fold factor. In another embodiment, translation is enhanced by a 10-fold factor. In another embodiment, translation is enhanced by a 15-fold factor. In another embodiment, translation is enhanced by a 20-fold factor. In another embodiment, translation is enhanced by a 50-fold factor. In another embodiment, translation is enhanced by a 100-fold factor. In another embodiment, translation is enhanced by a 200-fold factor. In another embodiment, translation is enhanced by a 500-fold factor. In another embodiment, translation is enhanced by a 1000-fold factor. In another embodiment, translation is enhanced by a 2000-fold factor. In another embodiment, the factor is 10-1000-fold. In another embodiment, the factor is 10-100-fold. In another embodiment, the factor is 10-200-fold. In another embodiment, the factor is 10-300-fold. In another embodiment, the factor is 10-500-fold. In another embodiment, the factor is 20-1000-fold. In another embodiment, the factor is 30-1000-fold. In another embodiment, the factor is 50-1000-fold. In another embodiment, the factor is 100-1000- fold. In another embodiment, the factor is 200-1000-fold. In another embodiment, translation is enhanced by any other significant amount or range of amounts. Each possibility represents a separate embodiment of the present disclosure.

[0390] Methods of determining translation efficiency are well known in the art, and include, e.g. measuring the activity of an encoded reporter protein (e.g., luciferase or renilla or green fluorescent protein [Wall A A, Phillips A M et al, Effective translation of the second cistron in two Drosophila dicistronic transcripts is determined by the absence of in-frame AUG codons in the first cistron. J Biol Chem 2005; 280(30): 27670-8]), or measuring radioactive label incorporated into the translated protein (Ngosuwan J, Wang N M et al., Roles of cytosolic Hsp70 and Hsp40Page 144 of 182 12197519v1molecular chaperones in post-translational translocation of pre-secretory proteins into the endoplasmic reticulum. J Biol Chem 2003; 278(9): 7034-42). Each method represents a separate embodiment of the present disclosure. Codon Optimization and GC Enrichment

[0391] The present disclosure also provides codon optimized nucleotide sequences.

[0392] As used herein, the term “codon-optimized” refers to alteration of codons in a coding region of a nucleic acid molecule (e.g., a nucleotide sequence) to reflect the typical codon usage of a host organism (e.g., a subject receiving a nucleic acid molecule (e.g., a nucleotide sequence)) preferably without altering the amino acid sequence encoded by the nucleic acid molecule. Within the context of the present disclosure, in some embodiments, coding regions are codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein. In some embodiments, codon-optimization may be performed such that codons for which frequently occurring tRNAs are available are inserted in place of “rare codons.” In some embodiments, codon-optimization may include increasing guanosine / cytosine (G / C) content of a coding region of RNA described herein as compared to the G / C content of the corresponding coding sequence of a wild type RNA, wherein the amino acid sequence encoded by the RNA is preferably not modified compared to the amino acid sequence.

[0393] In some embodiments, a coding sequence (also referred to as a “coding region”) is codon optimized for expression in the subject to whom a composition (e.g., a pharmaceutical composition) is to be administered (e.g., a human). Thus, in some embodiments, sequences in such a polynucleotide (e.g., a nucleotide sequence) may differ from wild type sequences encoding the relevant antigen, fragment or epitope thereof, even when the amino acid sequence of the antigen, fragment or epitope thereof is wild type.

[0394] In some embodiments, strategies for codon optimization for expression in a relevant subject (e.g., a human), and even, in some cases, for expression in a particular cell or tissue.

[0395] Various species exhibit particular bias for certain codons of a particular amino acid. Without wishing to be bound by any one theory, codon bias (differences in codon usage between organisms) often correlates with the efficiency of translation of an RNA, which is in turn believed to be dependent on, among other things, the properties of the codons being translated and the availability of particular transfer RNA (tRNA) molecules. The predominance of selected tRNAs in a cell may generally be a reflection of the codons used most frequently in peptide synthesis. Accordingly, genes may be tailored for optimal gene expression in a given organism based on codon optimization. Codon usage tables are available, for example, at the "Codon Usage Database" available at www.kazusa.orjp / codon / and these tables may be adapted in a number of ways. Computer algorithms for codon optimizing a particular sequence for expression in a particular subject or its cells are also available, such as Gene Forge (Aptagen; Jacobus, PA), are also available.

[0396] In some embodiments, a polynucleotide (e.g., a polyribonucleotide or a nucleotide sequence) of the present disclosure is codon optimized, wherein the codons in the polynucleotide (e.g., the polyribonucleotide) are adapted to human codon usage (herein referred to as “human codon optimized polynucleotide”). In some embodiments, a portion of a nucleotide sequence is codon optimized (e.g., a portion of or the portion encoding a glycoprotein or a portion of or the portion encoding a signal sequence). In some embodiments, the entire nucleotide sequence is codon optimized. Codons encoding the same amino acid occur at different frequencies in a subject, e.g.,Page 145 of 182 12197519v1a human. Accordingly, in some embodiments, the coding sequence of a polynucleotide of the present disclosure is modified such that the frequency of the codons encoding the same amino acid corresponds to the naturally occurring frequency of that codon according to the human codon usage, e.g., as shown in Table 5. For example, in the case of the amino acid Ala, the wild type coding sequence is preferably adapted in a way that the codon “GCC” is used with a frequency of 0.40, the codon “GCT” is used with a frequency of 0.28, the codon “GCA” is used with a frequency of 0.22 and the codon “GCG” is used with 30 a frequency of 0.10 etc. (see Table 5). Accordingly, in some embodiments, such a procedure (as exemplified for Ala) is applied for each amino acid encoded by the coding sequence of a polynucleotide to obtain sequences adapted to human codon usage. Table 5: Human codon usage with frequencies indicated for each amino acid. Amino Codon Frequency Amino Codon Frequency Acid Acid Al 1 P 11

[0397] Certain strategies for codon optimization and / or G / C enrichment for human expression are described in WO2002 / 098443, which is incorporated by reference herein in its entirety. In some embodiments, a coding sequence may be optimized using a multiparametric optimization strategy. In some embodiments, optimization parameters may include parameters that influence protein expression, which can be, for example, impacted on a transcriptionPage 146 of 182 12197519v1level, an RNA level, and / or a translational level. In some embodiments, exemplary optimization parameters include, but are not limited to transcription-level parameters (including, e.g., GC content, consensus splice sites, cryptic splice sites, SD sequences, TATA boxes, termination signals, artificial recombination sites, and combinations thereof); RNA- level parameters (including, e.g., RNA instability motifs, ribosomal entry sites, repetitive sequences, and combinations thereof); translation-level parameters (including, e.g., codon usage, premature poly(A) sites, ribosomal entry sites, secondary structures, and combinations thereof); or combinations thereof. In some embodiments, a coding sequence may be optimized by a GeneOptimizer algorithm as described in Fath et al. “Multiparameter RNA and Codon Optimization: A Standardized Tool to Assess and Enhance Autologous Mammalian Gene Expression” PLoS ONE 6(3): e17596; Rabb et al., which is incorporated herein by reference in its entirety, “The GeneOptimizer Algorithm: using a sliding window approach to cope with the vast sequence space in multiparameter DNA sequence optimization” Systems and Synthetic Biology (2010) 4:215-225; and Graft et al. “Codon-optimized genes that enable increased heterologous expression in mammalian cells and elicit efficient immune responses in mice after vaccination of naked DNA” Methods Mol Med (2004) 94:197-210, the entire content of each of which is incorporated herein for the purposes described herein. In some embodiments, a coding sequence may be optimized by Eurofins’ adaption and optimization algorithm “GENEius” as described in Eurofins’ Application Notes: Eurofins’ adaption and optimization software “GENEius” in comparison to other optimization algorithms, the entire content of which is incorporated by reference for the purposes described herein.

[0398] In some embodiments, a coding sequence utilized in accordance with the present disclosure has G / C content that is increased compared to a coding sequence for an HSV gC, gD, and / or gE (or immunogenic fragment thereof) construct described herein. In some embodiments, guanosine / cytidine (G / C) content of a coding region is modified relative to a comparable coding sequence for an HSV gC, gD, and / or gE (or immunogenic fragment thereof) construct described herein, but the amino acid sequence encoded by the nucleotide sequence is not modified.

[0399] Without wishing to be bound by any particular theory, it is proposed that GC enrichment may improve translation of a payload sequence. Typically, sequences having an increased G (guanosine) / C (cytidine) content are more stable than sequences having an increased A (adenosine) / U (uridine) content. In respect to the fact that several codons code for one and the same amino acid (so-called degeneration of the genetic code), the most favorable codons for the stability can be determined (so-called alternative codon usage). Depending on the amino acid to be encoded by a nucleotide sequence, there are various possibilities for modification of the ribonucleic acid sequence, compared to its wild type sequence. In particular, codons which contain A and / or U nucleosides can be modified by substituting these codons by other codons, which code for the same amino acids but contain no A and / or U or contain a lower content of A and / or U nucleosides.

[0400] In some embodiments, G / C content of a coding region of a nucleotide sequence described herein is increased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G / C content of the coding region prior to codon optimization, e.g., of the wild type RNA. In some embodiments, G / C content of a coding region of a nucleotide sequence described herein is decreased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G / C content of the coding region prior to codon optimization, e.g., of the wild type RNA.Page 147 of 182 12197519v1

[0401] In some embodiments, stability and translation efficiency of a nucleotide sequence may incorporate one or more elements established to contribute to stability and / or translation efficiency of the nucleotide sequence; exemplary such elements are described, for example, in PCT / EP2006 / 009448 incorporated herein by reference. In some embodiments, to increase expression of a nucleotide sequence used according to the present disclosure, a nucleotide sequence may be modified within the coding region, i.e., the sequence encoding the expressed peptide or protein, without altering the sequence of the expressed peptide or protein, for example so as to increase the GC- content to increase RNA stability and / or to perform a codon optimization and, thus, enhance translation in cells.

[0402] Compositions

[0403] In some embodiments, the present disclosure also provides a composition comprising a polyribonucleotide or RNA disclosed herein. In another embodiment, the present disclosure also provides a composition comprising an RNA construct disclosed herein.

[0404] In some embodiments, the present disclosure provides a composition comprising an RNA or a nucleoside- modified RNA encoding a polypeptide that comprises the glycoprotein B (gB) of HSV, or an immunogenic fragment thereof. In some embodiments, the composition comprises an RNA or a nucleoside-modified RNA encoding a polypeptide that comprises the gB of HSV-1, or an immunogenic fragment thereof. In another embodiment, the composition comprises an RNA or a nucleoside-modified RNA encoding a polypeptide that comprises the gB of HSV-2, or an immunogenic fragment thereof.

[0405] In some embodiments, the present disclosure describes compositions, which in some embodiments, are immunogenic compositions. In some embodiments, the immunogenic composition for use in the compositions and methods as described herein are vaccines. In some embodiments, RNA technology has the advantage that multiple immunogens can be included in a composition (e.g., immunogenic composition, e.g., a vaccine), rather than a more limited number of antigens using protein vaccines. Therefore, in some embodiments, a polyribonucleotide as described herein encodes one or more additional antigens or immunogens in addition to HSV-2 gB or portion thereof. In another embodiment, a composition as described herein comprises one or more additional polyribonucleotides encoding one or more additional antigens or immunogens in addition to the polyribonucleotide encoding HSV-2 gB or portion thereof.

[0406] In another embodiment, the additional antigens or immunogens comprise one or more HSV glycoproteins. In some embodiments, the HSV glycoprotein comprises glycoprotein H. In another embodiment, the HSV glycoprotein comprises glycoprotein L. In another embodiment, the HSV glycoprotein comprises glycoprotein I. In another embodiment, the HSV glycoprotein comprises glycoprotein C. In another embodiment, the HSV glycoprotein comprises glycoprotein D. In another embodiment, the HSV glycoprotein comprises glycoprotein E. In another embodiment, the one or more HSV glycoprotein comprises a combination of the glycoproteins described herein. In some embodiments, the additional antigens or immunogens are from HSV-1. In another embodiment, the additional antigens or immunogens are from HSV-2. Sequences for HSV-2 glycoprotein C, HSV-2 glycoprotein D and HSV-2 glycoprotein E have been published, see e.g. WO 2019 / 035066, which is incorporated herein by reference in its entirety.Page 148 of 182 12197519v1

[0407] In some embodiments, in addition to the polyribonucleotide encoding the HSV-2 gB disclosed herein, compositions disclosed herein further comprise RNA encoding one or more polypeptides encoded by immediate early genes such as ICP4, ICP10, and ICP27. In another embodiment, in addition to the polyribonucleotide encoding the HSV-2 gB disclosed herein, compositions disclosed herein further comprise RNA encoding one or more tegument structural proteins such as VP11 / 12, VP13 / 14, VP16 and VP22.

[0408] In another embodiment, a composition (e.g., immunogenic composition) disclosed herein, which in some embodiments is a vaccine, comprises one or more T cell immunogens. In another embodiment, a composition disclosed herein comprise immunogens from HSV-1 and HSV-2.

[0409] In some embodiments, the present disclosure provides a composition comprising (i) an RNA or a nucleoside-modified RNA encoding the HSV-2 gB disclosed herein, or an immunogenic fragment thereof, and (ii) RNA or a nucleoside-modified RNA encoding one or more HSV-2 glycoprotein H, glycoprotein L, and glycoprotein I, or an immunogenic fragment thereof.

[0410] In another embodiment, the present disclosure provides a composition comprising (i) an RNA or a nucleoside-modified RNA encoding the HSV-2 gB disclosed herein, or an immunogenic fragment thereof, and (ii) RNA or a nucleoside-modified RNA encoding HSV-2 glycoproteins C, D and E, or an immunogenic fragment thereof. Sequences for HSV-2 glycoproteins C, D and E have been published, see e.g. WO 2019 / 035066, which is incorporated herein by reference in its entirety. In other embodiments, the RNA or nucleoside-modified RNA encoding HSV-2 glycoprotein C or an immunogenic fragment thereof comprises one or more sequences as set forth in SEQ ID NOs: 259 or 265. In other embodiments, the RNA or nucleoside-modified RNA encoding HSV-2 glycoprotein D or an immunogenic fragment thereof comprises one or more sequences as set forth in SEQ ID NOs: 261 or 267. In other embodiments, the RNA or nucleoside-modified RNA encoding HSV-2 glycoprotein E or an immunogenic fragment thereof comprises one or more sequences as set forth in SEQ ID NOs: 263 or 269.

[0411] In other embodiments, the RNA or nucleoside-modified RNA encoding HSV-2 glycoprotein C or an immunogenic fragment thereof encodes a polypeptide as set forth in SEQ ID NOs: 258 or 264. In other embodiments, the RNA or nucleoside-modified RNA encoding HSV-2 glycoprotein D or an immunogenic fragment thereof encodes a polypeptide as set forth in SEQ ID NOs: 260 or 266. In other embodiments, the RNA or nucleoside-modified RNA encoding HSV-2 glycoprotein E or an immunogenic fragment thereof encodes a polypeptide as set forth in SEQ ID NOs: 262 or 268.

[0412] In some embodiments, the present disclosure provides a composition comprising (i) an RNA or a nucleoside-modified RNA encoding the HSV-2 gB disclosed herein, or an immunogenic fragment thereof, and (ii) RNA encoding one or more polypeptides encoded by immediate early genes such as ICP0, ICP4, ICP10, and ICP27, or an immunogenic fragment thereof. In some embodiments, the present disclosure provides a composition comprising (i) an RNA or a nucleoside-modified RNA encoding the HSV-2 gB disclosed herein, or an immunogenic fragment thereof, (ii) an RNA or a nucleoside-modified RNA encoding ICP0, or an immunogenic fragment thereof, and (iii) an RNA or a nucleoside-modified RNA encoding ICP4, or an immunogenic fragment thereof. In another embodiment, the composition comprises (i) an RNA or a nucleoside-modified RNA encoding the HSV-2 gB disclosed herein, or an immunogenic fragment thereof, and (ii) RNA encoding one or more tegument structural proteins such as VP11 / 12, VP13 / 14, VP16 and VP22, or an immunogenic fragment thereof.Page 149 of 182 12197519v1

[0413] In some embodiments, the composition comprising an RNA or a nucleoside-modified RNA encoding the HSV-2 gB disclosed herein, or an immunogenic fragment thereof, further comprises one or more RNAs or nucleoside- modified RNAs encoding (a) HSV glycoprotein C (gC) or immunogenic fragment thereof, (b) HSV glycoprotein D (gD) or immunogenic fragment thereof, (c) HSV glycoprotein E (gE) or immunogenic fragment thereof, (d) HSV glycoprotein H (gH) or immunogenic fragment thereof, (e) HSV glycoprotein L (gL) or immunogenic fragment thereof, (f) HSV glycoprotein I (gI) or immunogenic fragment thereof, or (g) any combination thereof.

[0414] In some embodiments, the composition comprising an RNA or a nucleoside-modified RNA encoding the HSV-2 gB disclosed herein, or an immunogenic fragment thereof, further comprises a nanoparticle, a lipid, a polymer, cholesterol, or a cell penetrating peptide. In some embodiments, the nanoparticle can be a lipid nanoparticle. In some embodiments, the nanoparticle, lipid, polymer, cholesterol, or cell penetrating peptide encapsulates the polyribonucleotide or RNA construct disclosed herein.

[0415] In some embodiments, the one or more polyribonucleotides are encapsulated in a nanoparticle. In some embodiments, the one or more polyribonucleotides are encapsulated in a lipid. In some embodiments, the one or more polyribonucleotides are encapsulated in a polymer. In some embodiments, the one or more polyribonucleotides are encapsulated in a cholesterol. In some embodiments, the one or more polyribonucleotides are encapsulated in a cell penetrating peptide.

[0416] In some embodiments, the one or more polyribonucleotides are attached to a nanoparticle. In some embodiments, the one or more polyribonucleotides are attached to a lipid. In some embodiments, the one or more polyribonucleotides are attached to a polymer. In some embodiments, the one or more polyribonucleotides are attached to a cholesterol. In some embodiments, the one or more polyribonucleotides are attached to a cell penetrating peptide.

[0417] In some embodiments, a method of present disclosure further comprises mixing the RNA with a transfection reagent prior to the step of contacting. In another embodiment, a method of present disclosure further comprises administering the RNA together with the transfection reagent. In another embodiment, the transfection reagent is a cationic lipid reagent.

[0418] In another embodiment, the transfection reagent is a lipid-based transfection reagent. In another embodiment, the transfection reagent is a protein-based transfection reagent. In another embodiment, the transfection reagent is a polyethyleneimine based transfection reagent. In another embodiment, the transfection reagent is calcium phosphate. In another embodiment, the transfection reagent is Lipofectin® or Lipofectamine®. In another embodiment, the transfection reagent is any other transfection reagent known in the art.

[0419] In another embodiment, the transfection reagent forms a liposome. Liposomes, in another embodiment, increase intracellular stability, increase uptake efficiency and improve biological activity.

[0420] In another embodiment, liposomes are hollow spherical vesicles composed of lipids arranged in a similar fashion as those lipids which make up the cell membrane. They have, in another embodiment, an internal aqueous space for entrapping water soluble compounds and range in size from 0.05 to several microns in diameter. In another embodiment, liposomes can deliver RNA to cells in a biologically active form (see Langer, Science 249:1527-Page 150 of 182 12197519v11...

Claims

CLAIMS What is claimed is:

1. A nucleoside-modified polyribonucleotide encoding a polypeptide, wherein said polypeptide comprises the ectodomain of a Herpes Simplex Virus-2 (HSV-2) glycoprotein B (gB) antigen having the amino acid sequence as set forth in SEQ ID NO:

1.

2. The polyribonucleotide of claim 1, wherein the nucleoside modification comprises one or more pseudouridine residues.

3. The polyribonucleotide of claim 2, wherein said one or more pseudouridine residues comprise m1Ψ (1- methylpseudouridine).

4. The polyribonucleotide of claim 2, wherein said one or more pseudouridine residues comprise m1acp3Ψ (1- methyl-3-(3-amino-5-carboxypropyl)pseudouridine, Ψm (2′-O-methylpseudouridine), m5D (5- methyldihydrouridine), m3Ψ (3-methylpseudouridine), or any combination thereof.

5. The polyribonucleotide of any one of claims 1-4, further comprising a secretory signal sequence.

6. The polyribonucleotide of any one of claims 1-5, comprising the ribonucleic acid sequence as set forth in SEQ ID NO:

5.

7. The polyribonucleotide of claim 5, wherein said secretory signal sequence encodes an IL2 secretory signal polypeptide.

8. The polyribonucleotide of claim 7, wherein said IL2 secretory signal polypeptide comprises the amino acid sequence as set forth in SEQ ID NO:

2.

9. The polyribonucleotide of claim 7, wherein said IL2 secretory signal polypeptide is encoded by the ribonucleic acid sequence as set forth in SEQ ID NO:

3.

10. The polyribonucleotide of claim 5, wherein said secretory signal sequence encodes an HSV gB secretory signal polypeptide.

11. The polyribonucleotide of claim 10, wherein the gB secretory signal polypeptide comprises the amino acid sequence as set forth in SEQ ID NO:

4.

12. The polyribonucleotide of any one of claims 1-11, wherein said polyribonucleotide further comprises a 5′ untranslated region.

13. The polyribonucleotide of any one of claims 1-12, wherein said polyribonucleotide further comprises a cap- independent translational enhancer.

14. The polyribonucleotide of any one of claims 1-13, wherein said polyribonucleotide further comprises a 3′ untranslated region.Page 179 of 182 12197519v115. The polyribonucleotide of any one of claims 1-14, wherein said polyribonucleotide further comprises a poly-A tail.

16. The polyribonucleotide of any one of claims 1-15, wherein said polyribonucleotide further comprises an m7GpppG cap, 3′-O-methyl-m7GpppG cap, or anti-reverse cap analog.

17. The polyribonucleotide of any one of claims 1-9 or 12-16, comprising the ribonucleic acid sequence as set forth in SEQ ID NO:

6.

18. A composition or combination comprising the polyribonucleotide of any one of claims 1-17.

19. The composition or combination of claim 18, wherein said composition comprises a nanoparticle, lipid, polymer, cholesterol, or cell penetrating peptide.

20. The composition or combination of claim 19, wherein said nanoparticle, lipid, polymer, cholesterol, or cell penetrating peptide encapsulates a polyribonucleotide.

21. The composition or combination of claim 20, wherein said nanoparticle is a lipid nanoparticle.

22. The composition or combination of any one of claims 18-21, further comprising one or more RNAs encoding a) HSV glycoprotein C (gC) or immunogenic fragment thereof, b) HSV glycoprotein D (gD) or immunogenic fragment thereof, c) HSV glycoprotein E (gE) or immunogenic fragment thereof, d) HSV glycoprotein H (gH) or immunogenic fragment thereof, e) HSV glycoprotein L (gL) or immunogenic fragment thereof, f) HSV glycoprotein I (gI) or immunogenic fragment thereof, or g) any combination thereof.

23. A polypeptide encoded by the polyribonucleotide of any one of claims 1-17.

24. A host cell comprising the polyribonucleotide of any one of claims 1-17.

25. A host cell comprising a polypeptide of claim 23.

26. A method of treating a Herpes Simplex Virus (HSV) infection in a subject, comprising the step of administering to said subject the polyribonucleotide of any one of claims 1-17 or the composition or combination of any one of claims 18-22.

27. A method of suppressing, inhibiting, or reducing the incidence of a Herpes Simplex Virus (HSV) infection in a subject, comprising the step of administering to said subject the polyribonucleotide of any one of claims 1-17 or the composition or combination of any one of claims 18-22.

28. The method of claim 26 or 27, wherein said HSV infection comprises an HSV-1 infection.

29. The method of claim 26 or 27, wherein said HSV infection comprises an HSV-2 infection.Page 180 of 182 12197519v130. The method of claim 26 or 27, wherein said HSV infection is a primary infection or a secondary infection.

31. The method of claim 26 or 27, wherein said HSV infection comprises a flare, recurrence, HSV labialis following a primary HSV infection, a reactivation of a latent HSV infection, an HSV encephalitis, an HSV neonatal infection, a genital HSV infection, an oral HSV infection, or a combination thereof.

32. The method of claim 26 or 27, wherein said HSV infection is a latent HSV infection.

33. The method of claim 32, wherein said latent HSV infection comprises a genital HSV infection or an oral HSV infection.

34. The method of claim 26 or 27, wherein said administration comprises intramuscular, subcutaneous, intradermal, intranasal, intravaginal, intrarectal, or topical administration.

35. A method of inducing an anti-HSV immune response in a subject, comprising the step of administering to said subject the polyribonucleotide of any one of claims 1-17 or the composition or combination of any one of claims 18-22.

36. The method of claim 35, wherein said immune response comprises a CD4 immune response, a CD8 immune response, a T follicular helper cell immune response, a germinal center B cell immune response, an IgG antibody response to gB2, or a combination thereof.

37. The method of claim 35, wherein said administration comprises intramuscular, subcutaneous, intradermal, intranasal, intravaginal, intrarectal, or topical administration.Page 181 of 182 12197519v1

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