Mutant fragment of VZV glycoprotein E

Abstract

Provided herein are fragments of the gE protein of varicella-zoster virus and therapeutic nucleic acid molecules for the management, prevention, and / or treatment of diseases or disorders caused by varicella-zoster virus or infection thereof. Also provided herein are therapeutic compositions comprising vaccines and lipid nanoparticles containing therapeutic nucleic acids, as well as related therapeutic methods and uses.

Description

[Technical Field]

[0001] 1. Field This disclosure relates, in general, to fragments of varicella-zoster virus (VZV) glycoprotein E (gE) and nucleic acid molecules that can be used for the management, prevention, and treatment of diseases or disorders caused by VZV or infection with VZV. This disclosure also relates to lipid-containing compositions of nucleic acid molecules (including vaccines). [Background technology]

[0002] 2.Background Varicella-zoster virus (VZV), also known as human herpesvirus type 3, is a double-stranded DNA virus belonging to the alphaherpesvirus class. VZV has only one serotype. The VZV genome contains 71 genes and encodes 67 different proteins, including six glycoproteins currently named gE, gB, gH, gI, gC, and gL. Glycoproteins gE, gB, and gH are very abundant in infected cells and are also present in the virion envelope. Antibodies induced by three major glycoproteins can neutralize the virus. Specific humoral and cellular immunity, as well as cytokines such as interferons, play a significant role in suppressing and restoring VZV spread, with specific cellular immunity being particularly important.

[0003] Zostabax (MSD) is a weakened viral vaccine that can reduce the burden by 61.1% (65.5% for those aged 60-69 and 55.4% for those aged 70 and over). It can also reduce the duration of pain and discomfort caused by the virus. Shinrix (GSK) is a subunit vaccine containing gE and the adjuvant system AS01B, which enhances the cellular immune response. Its efficacy can reach over 90%. [Overview of the Initiative]

[0004] 4. Overview In one embodiment, provided herein are fragments of the gE protein of VZV, which optionally contain mutations (e.g., substitutions).

[0005] In some embodiments, the fragment contains a cleavage of at least one amino acid residue and up to 49 amino acid residues from the C-terminus compared to a mature gE protein. In some embodiments, the fragment contains a cleavage of 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or one(s) amino acid residues from the C-terminus compared to a mature gE protein. In some embodiments, the fragment contains cleavage of 11–18 (e.g., 11, 12, 13, 14, 15, 16, 17, or 18) or 34–44 (e.g., 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44) amino acid residues from the C-terminus compared to a mature gE protein. In some embodiments, the fragment contains cleavage of 12–16 (e.g., 12, 13, 14, 15, or 16) or 35–39 (e.g., 35, 36, 37, 38, or 39) amino acid residues from the C-terminus compared to a mature gE protein. In some embodiments, the fragment contains cleavage of 14 or 37 amino acid residues from the C-terminus compared to a mature gE protein.

[0006] In some embodiments, the fragment includes substitution Y569A. In some embodiments, the fragment includes substitution Y582G. In some embodiments, the fragment includes substitutions Y569A and Y582G. In some embodiments, the fragment includes substitution S593A. In some embodiments, the fragment includes substitution S595A. In some embodiments, the fragment includes substitution T596A. In some embodiments, the fragment includes substitution T598A. In some embodiments, the fragment includes substitutions S593A, S595A, T596A, and T598A. In some embodiments, the fragment includes substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A. In such embodiments, the amino acid positions are numbered based on the full-length gE protein.

[0007] In some embodiments, the fragment contains cleavage of 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus compared to the mature gE protein. In such embodiments, the fragment optionally contains substitution Y569A.

[0008] In some embodiments, the fragment contains cleavage of 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, or 31 amino acid residues from the C-terminus compared to a mature gE protein. In such embodiments, the fragment optionally contains substitution Y582G, with or without substitution Y569A. In such embodiments, the fragment contains substitution Y569A and Y582G.

[0009] In some embodiments, the fragment includes a cleavage of 30 or 29 amino acid residues from the C-terminus compared to a mature gE protein. In such embodiments, the fragment optionally includes substitution S593A and is accompanied or absent with substitution Y569A and / or substitution Y582G. In such embodiments, the fragment includes substitution Y569A, Y582G, and S593A. In some embodiments, the fragment includes a cleavage of 28 amino acid residues from the C-terminus compared to a mature gE protein. In such embodiments, the fragment optionally includes substitution S595A and is accompanied or absent with substitution Y569A and / or Y582G and / or S593A. In such embodiments, the fragment includes substitution Y569A, Y582G, S593A, and S595A. In some embodiments, the fragment includes a cleavage of 27 or 26 amino acid residues from the C-terminus compared to a mature gE protein. In such embodiments, the fragment optionally contains substituted T596A and is accompanied or absent substituted Y569A and / or Y582G and / or S593A and / or S595A. In such embodiments, the fragment contains substituted Y569A, Y582G, S593A, S595A, and T596A. In some embodiments, the fragment, compared to a mature gE protein, contains cleavage of 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 amino acid residues, or 1 amino acid residue. In such embodiments, the fragment optionally contains substituted T598A and is accompanied or absent substituted Y569A and / or Y582G and / or S593A and / or S595A and / or T596A. In such embodiments, the fragment includes substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A.

[0010] In some embodiments, the mature gE protein contains the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length gE protein contains the amino acid sequence described in SEQ ID NO: 55.

[0011] In some embodiments, the fragment includes an amino acid sequence described in SEQ ID NOs: 3, 6, 8, 10, or 12, or an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence described in SEQ ID NOs: 3, 6, 8, 10, or 12.

[0012] In some embodiments, the N-terminus of the fragment is fused to the C-terminus of the signal peptide. In some embodiments, the N-terminus of the fragment is fused to the native signal peptide of the VZV gE protein. In some embodiments, the native signal peptide includes the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the N-terminus of the fragment is fused to the C-terminus of the human tPA signal peptide. In some embodiments, the human tPA signal peptide includes the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the N-terminus of the fragment is fused to the C-terminus of the human IgE signal peptide. In some embodiments, the human IgE signal peptide includes the amino acid sequence described in SEQ ID NO: 23.

[0013] In one embodiment, what is provided herein is a nucleic acid encoding a fragment described herein.

[0014] In some embodiments, the fragment is encoded by a nucleotide sequence described in SEQ ID NOs: 4, 5, 7, 9, 11, or 13, or by a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the nucleotide sequence described in SEQ ID NOs: 4, 5, 7, 9, 11, or 13.

[0015] In some embodiments, the native signal peptide of the VZV gE protein is encoded by a nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22, or a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22. In some embodiments, the human tPA signal peptide is encoded by a nucleotide sequence described in SEQ ID NO: 28, or a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the nucleotide sequence described in SEQ ID NO: 28. In some embodiments, the human IgE signal peptide is encoded by a nucleotide sequence described in SEQ ID NO: 24, 25, or 26, or a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the nucleotide sequence described in SEQ ID NO: 24, 25, or 26.

[0016] In some embodiments, the proteins provided herein include variants of the mature glycoprotein E (gE) of varicella-zoster virus (VZV), the variants being (a) (i) cleavage of 37 amino acid residues from the C-terminus of mature gE, and (ii) amino acid residue substitutions Y569A and Y582G (amino acid residue positions 569 and 582 are amino acid residue position numbers of full-length VSV gE); (b) amino acid residue substitutions Y569A and Y582G (amino acid residue positions 569 and 582 are amino acid residue position numbers of full-length VSV gE); (c) amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid residue positions 569, 582, 593, 595, 596, and 598 are full-length VSV (d) Amino acid residue position numbers of gE; (i) (i) cleavage of 50 amino acid residues from the C-terminus of a mature gE protein, and (ii) amino acid residue substitution Y569A (amino acid residue position 569 is the amino acid residue position number of full-length VSV gE). In some embodiments, the mutant includes the amino acid sequence of SEQ ID NO: 6, 8, 10, 12, or 3. In some embodiments, the mutant consists of the amino acid sequence of SEQ ID NO: 6, 8, 10, 12, or 3. In some embodiments, the mutant includes the amino acid sequence of SEQ ID NO: 6. In some embodiments, the mutant consists of the amino acid sequence of SEQ ID NO: 6. In some embodiments, the variant includes an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95% identical to SEQ ID NOs. 6, 8, 10, or 12. In some embodiments, the variant includes an amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NOs. 6, 8, 10, or 12.In some embodiments, the variant contains an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95% identical to SEQ ID NO: 6. In some embodiments, the variant contains an amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 6. In some embodiments, the mature gE contains the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE contains the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the mature gE contains the amino acid sequence described in SEQ ID NO: 1, and the full-length VZV gE contains the amino acid sequence described in SEQ ID NO: 55.

[0017] In some embodiments, the protein further comprises a VZV gE signal peptide. In some embodiments, the VZV gE signal peptide comprises the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the VZV gE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the protein further comprises a heterologous signal peptide, the N-terminus of the variant fused to the C-terminus of the heterologous signal peptide. In some embodiments, the heterologous signal peptide is a human tPA signal peptide. In some embodiments, the human tPA signal peptide comprises the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the human tPA signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the amino acid sequence of the human tPA signal peptide consists of the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the heterologous signal peptide is a human IgE signal peptide. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the protein comprises the amino acid sequence described in SEQ ID NO: 59.

[0018] In some embodiments, the herein provides a protein comprising a variant of mature VZV gE and a human IgE signal peptide, wherein the variant comprises the amino acid sequence described in SEQ ID NO: 6, and the amino acid sequence of the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23.

[0019] In some embodiments, what is provided herein is a protein comprising the amino acid sequence described in SEQ ID NO: 59. In some embodiments, what is provided herein is a protein comprising an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to the amino acid sequence described in SEQ ID NO: 59. In some embodiments, what is provided herein is a protein comprising an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 59. In some embodiments, what is provided herein is a protein whose amino acid sequence consists of the amino acid sequence described in SEQ ID NO: 59.

[0020] In some embodiments, what is provided herein is a fragment of mature gE of VZV, the fragment comprising a cleavage of at least one amino acid residue, up to 50, 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage comprises a cleavage of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of 11, 12, 13, 14, 15, 16, 17, 18, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of 14 or 37 amino acid residues from the C-terminus of mature gE. In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions. In some embodiments, the fragment further comprises one, two, three, four, five, or all of the following amino acid substitutions: Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid position numbers according to full-length VZV gE). In some embodiments, the fragment further comprises the amino acid residue substitution Y569A, where amino acid residue position 569 is an amino acid residue position number according to full-length VZV gE. In some embodiments, the fragment includes an amino acid sequence that is 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%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment includes an amino acid sequence that is at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, or at least 85% identical to the amino acid sequence described in SEQ ID NO: 6, 3, 8, 10, or 12.In some embodiments, the fragment includes an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment includes the amino acid sequence described in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the amino acid sequence of the fragment consists of the amino acid sequence described in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment includes the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the amino acid sequence of the fragment consists of the amino acid sequence described in SEQ ID NO: 6.

[0021] In some embodiments, what is provided herein is a fragment of mature gE of VZV, the fragment comprising a cleavage of up to 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, or 31 amino acid residues from the C-terminus of mature gE. In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions. In some embodiments, the fragment further comprises one, two, three, four, five, or all of the following amino acid substitutions: Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid position numbers according to full-length VZV gE). In some embodiments, the fragment further comprises the amino acid residue substitution Y582G, where amino acid residue position 582 is the amino acid residue position number according to the full-length VZV gE protein. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, mature gE contains the amino acid sequence described in SEQ ID NO: 1, and full-length VZV gE contains the amino acid sequence described in SEQ ID NO: 55.

[0022] In some embodiments, what is provided herein is a fragment of mature gE of VZV, the fragment comprising a cleavage of up to 30 or 29 amino acid residues from the C-terminus of mature gE. In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions. In some embodiments, the fragment further comprises one, two, three, four, five, or all of the following amino acid substitutions: Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid position numbers according to full-length VZV gE). In some embodiments, the fragment further comprises the amino acid residue substitution S593A, where amino acid residue position 593 is an amino acid residue position number according to full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1, and the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55.

[0023] In some embodiments, what is provided herein is a fragment of mature gE of VZV, the fragment comprising a cleavage of up to 28 amino acid residues from the C-terminus of mature gE. In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions. In some embodiments, the fragment further comprises one, two, three, four, five, or all of the following amino acid substitutions: Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid position numbers according to full-length VZV gE). In some embodiments, the fragment comprises the amino acid residue substitution S595A, where amino acid residue position 595 is an amino acid residue position number according to full-length VSV gE. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1, and the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55.

[0024] In some embodiments, what is provided herein is a fragment of mature gE of VZV, the fragment comprising a cleavage of up to 27 or 26 amino acid residues from the C-terminus of mature gE. In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions. In some embodiments, the fragment further comprises one, two, three, four, five, or all of the following amino acid substitutions: Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid position numbers according to full-length VZV gE). In some embodiments, the fragment further comprises the amino acid residue substitution T596A, where amino acid residue position 596 is an amino acid residue position number according to full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1, and the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55.

[0025] In some embodiments, provided herein are fragments of mature gE of VZV, the fragment comprising up to 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 amino acid residues, or up to 1 amino acid residue, from the C-terminus of mature gE. In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions. In some embodiments, the fragment further comprises one, two, three, four, five or all of the following amino acid substitutions: Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid position numbers according to full-length VZV gE). In some embodiments, the fragment further comprises the amino acid residue substitution T598A, where amino acid residue position 598 is an amino acid residue position number according to full-length VZV gE. In some embodiments, mature gE contains the amino acid sequence described in SEQ ID NO: 1. In some embodiments, full-length VZV gE contains the amino acid sequence described in SEQ ID NO: 55. In some embodiments, mature gE contains the amino acid sequence described in SEQ ID NO: 1, and full-length VZV gE contains the amino acid sequence described in SEQ ID NO: 55.

[0026] In some embodiments, provided herein are fragments of mature gE of VZV, the fragment comprising a cleavage of 37 amino acid residues from the C-terminus of mature gE. In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions. In some embodiments, the fragment further comprises one, two, three, four, five, or all of the following amino acid substitutions: Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid position numbers according to full-length VZV gE). In some embodiments, the fragment comprises (1) a cleavage of 37 amino acid residues from the C-terminus of mature gE, and (2) amino acid residue substitutions Y569A and Y582G, where amino acid residue positions 569 and 582 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, mature gE contains the amino acid sequence described in SEQ ID NO: 1, and full-length VZV gE contains the amino acid sequence described in SEQ ID NO: 55.

[0027] In some embodiments, provided herein is a fusion protein comprising a fragment of mature VZV gE and a heterologous signal peptide, wherein the N-terminus of the fragment is fused to the C-terminus of the heterologous signal peptide. In some embodiments, the heterologous signal peptide is a human tPA signal peptide. In some embodiments, the human tPA signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 28. In some embodiments, the human tPA signal peptide comprises the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the heterologous signal peptide is a human IgE signal peptide. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the fragment includes a cleavage of at least one amino acid residue, up to 50, 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is a cleavage of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of 11, 12, 13, 14, 15, 16, 17, 18, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of 14 or 37 amino acid residues from the C-terminus of mature gE. In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions.In some embodiments, the fragment further comprises one, two, three, four, five, or all of the following amino acid substitutions: Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid position numbers correspond to full-length VZV gE). In some embodiments, the fragment is the fragment described herein. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1, and the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55.

[0028] In some embodiments, provided herein is a fusion protein comprising a fragment of mature VZV gE and a human IgE signal peptide, wherein the fragment comprises a cleavage of 37 amino acid residues from the C-terminus of mature gE, and the N-terminus of the fragment is fused to the C-terminus of the human IgE signal peptide. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1, and the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55.

[0029] In some embodiments, the herein provides a fusion protein comprising a fragment of mature VZV gE and a human IgE signal peptide, wherein the fragment comprises the amino acid sequence described in SEQ ID NO: 6, and the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23, with the N-terminus of the fragment fused to the C-terminus of the human IgE signal peptide. In some embodiments, the amino acid sequence of the fragment consists of the amino acid sequence described in SEQ ID NO: 6, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the herein provides a fusion protein comprising a fragment of mature VZV gE and a human IgE signal peptide, wherein the amino acid sequence of the fragment consists of the amino acid sequence described in SEQ ID NO: 6, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23, with the N-terminus of the fragment fused to the C-terminus of the human IgE signal peptide.

[0030] In some embodiments, what is provided herein is a nucleic acid encoding a protein described herein. In some embodiments, what is provided herein is a nucleic acid encoding a fragment described herein. In some embodiments, what is provided herein is a nucleic acid encoding a fusion protein described herein. In some embodiments, what is provided herein is a nucleic acid comprising a protein described herein, a fragment described herein, or a fusion protein described herein, wherein the nucleic acid comprises a nucleotide sequence described in SEQ ID NOs: 7, 9, 11, 13, 4, or 5. In some embodiments, what is provided herein is a nucleic acid comprising a protein described herein, a fragment described herein, or a fusion protein described herein, wherein the nucleic acid has at least 80%, at least 85%, at least 90%, at least 91%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with respect to the nucleotide sequence described in SEQ ID NOs: 7, 9, 11, 13, 4, or 5. In some embodiments, provided herein is a nucleic acid comprising a protein, a fragment, or a fusion protein as described herein, wherein the nucleic acid comprises a nucleotide sequence that is at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identical to the nucleotide sequence described in SEQ ID NO: 7, 9, 11, 13, 4, or 5. In some embodiments, provided herein is a nucleic acid comprising a protein, a fragment, or a fusion protein as described herein, wherein the nucleic acid comprises a nucleotide sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to the nucleotide sequence described in SEQ ID NO: 7, 9, 11, 13, 4, or 5.In some embodiments, what is provided herein is a nucleic acid comprising a protein, a fragment, or a fusion protein as described herein, wherein the nucleic acid comprises a nucleotide sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the nucleotide sequence described in SEQ ID NO: 7, 9, 11, 13, 4, or 5. In some embodiments, what is provided herein is a nucleic acid comprising a protein, a fragment, or a fusion protein as described herein, wherein the nucleic acid comprises a nucleotide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 7, 9, 11, 13, 4, or 5. In some embodiments, what is provided herein is a nucleic acid comprising a protein, a fragment, or a fusion protein as described herein, wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 7.

[0031] In some embodiments, provided herein is a nucleic acid encoding a protein described herein, the protein comprising a variant of mature gE of VZV and a human IgE signal peptide, the nucleotide sequence encoding the IgE signal peptide comprising the nucleotide sequence described in SEQ ID NO: 24, 25, or 26. In some embodiments, provided herein is a nucleic acid encoding a protein described herein, the protein comprising a variant of mature gE of VZV and a human IgE signal peptide, the nucleotide sequence encoding the IgE signal peptide comprising a nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 24, 25, or 26.

[0032] In some embodiments, provided herein is a nucleic acid encoding a fusion protein as described herein, the fusion protein comprising a fragment of mature gE from VZV and a human IgE signal peptide, wherein the nucleotide sequence encoding the IgE signal peptide comprises the nucleotide sequence described in SEQ ID NO: 24, 25, or 26.

[0033] In some embodiments, provided herein is a nucleic acid encoding a protein described herein, the protein comprising a variant of mature gE of VZV and a VZV gE signal peptide, the nucleotide sequence of the signal peptide comprising the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22. In some embodiments, provided herein is a nucleic acid encoding a protein described herein, the protein comprising a variant of mature gE of VZV and a VZV gE signal peptide, the nucleotide sequence of the signal peptide comprising a nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22.

[0034] In some embodiments, provided herein is a nucleic acid encoding a protein described herein, the protein comprising a variant of mature gE of VZV and a human tPA signal peptide, the nucleotide sequence encoding the human tPA signal peptide comprising the nucleotide sequence described in SEQ ID NO: 28. In some embodiments, provided herein is a nucleic acid encoding a protein described herein, the protein comprising a fragment of mature gE of VZV and a human tPA signal peptide, the nucleotide sequence encoding the human tPA signal peptide comprising a nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 28.

[0035] In some embodiments, provided herein is a nucleic acid encoding the fusion protein described herein, the fusion protein comprising a fragment of mature gE of VZV and a human tPA signal peptide, wherein the nucleotide sequence encoding the human tPA signal peptide comprises the nucleotide sequence described in SEQ ID NO: 28. In some embodiments, provided herein is a nucleic acid encoding the fusion protein described herein, the fusion protein comprising a fragment of mature gE of VZV and a human tPA signal peptide, wherein the nucleotide sequence encoding the human tPA signal peptide comprises a nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 28.

[0036] In some embodiments, disclosed herein is a vector or cells comprising the nucleic acid described herein. In some embodiments, the vector is preferably an IVT plasmid. In some embodiments, disclosed herein is a composition comprising the fragment or nucleic acid described herein. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the composition is a vaccine.

[0037] In one embodiment, provided herein are non-naturally occurring nucleic acid molecules that can be used for the prevention, management, and treatment of diseases or disorders caused by or resulting from VZV.

[0038] In some embodiments, nucleic acids not found in nature include coding nucleotide sequences encoding the fragments described herein. In some embodiments, the fragment consists of, essentially, or includes an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence described in SEQ ID NO: 1 or the fragment thereof. In some embodiments, the fragment consists of, essentially, or includes an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence described in SEQ ID NO: 3, 6, 8, 10, or 12. In some embodiments, the fragment consists of, essentially, or includes an amino acid sequence described in SEQ ID NO: 3, 6, 8, 10, or 12. In some embodiments, the coding nucleotide sequence consists of, essentially, or includes, a nucleotide sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the nucleotide sequence or fragment described in SEQ ID NO: 2. In some embodiments, the coding nucleotide sequence is codon-optimized for expression in the target cell. In some embodiments, the target is a non-human mammal. In some embodiments, the target is human. In some embodiments, the coding nucleotide sequence consists of, essentially, or includes a nucleotide sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the nucleotide sequence described in SEQ ID NOs: 4, 5, 7, 9, 11, or 13. In some embodiments, the fragment is fused to a gE native signal peptide.In some embodiments, the signal peptide consists of, or comprises, an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the signal peptide is encoded by a coding nucleotide sequence consisting of, or comprising, a nucleotide sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with respect to the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22. In some embodiments, the fragment is fused to a heterologous polypeptide. In some embodiments, the heterologous polypeptide is selected from the Fc region of human immunoglobulin, signal peptides, and peptides that promote the polymerization of the fusion protein. In some embodiments, the signal peptide is a signal peptide derived from IgE or tPA. In some embodiments, the signal peptide consists of, or comprises, an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the signal peptide is encoded by a coding nucleotide sequence that is essentially or includes a nucleotide sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the nucleotide sequence described in SEQ ID NO: 24, 25, or 26. In some embodiments, the signal peptide is essentially or includes an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence described in SEQ ID NO: 27.In some embodiments, the signal peptide is encoded by a coding nucleotide sequence that is essentially derived from, or contains, a nucleotide sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with respect to the nucleotide sequence described in SEQ ID NO: 28. In some embodiments, the multimerization is dimerization or trimerization. In some embodiments, the nucleic acid that does not exist in nature further comprises a 5' untranslated region (5'-UTR), the 5'-UTR comprising the sequence described in any one of SEQ ID NOs: 29-38. In some embodiments, the nucleic acid that does not exist in nature further comprises a 3' untranslated region (3'-UTR), the 3'-UTR comprising the sequence described in any one of SEQ ID NOs: 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal. In some embodiments, the nucleic acid comprises a nucleotide sequence described in SEQ ID NOs. 49, 50, 51, 52, 53, 54, 60, 61, 62, 63, or 64, or a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a nucleotide sequence described in SEQ ID NOs. 49, 50, 51, 52, 53, 54, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid, which does not exist in nature, comprises one or more functional nucleotide analogs. In some embodiments, the nucleic acid, which does not exist in nature, comprises one or more functional nucleotide analogs selected from pseudouridine (psd), 1-methyl-psoiduridine (m1), and 5-methylcytosine. In some embodiments, the nucleic acid is DNA or mRNA.In some embodiments, the nucleic acid comprises, is essentially, or includes the nucleotide sequence described in SEQ ID NO: 63, or comprises a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the nucleotide sequence described in SEQ ID NO: 63, wherein all thymine (T) is substituted with uracil (U) or N1-methylpsoiduridine, and / or the first nucleotide G is m. 7 It has been replaced with GpppAmpU.

[0039] In some embodiments, disclosed herein are vectors or cells containing the nucleic acid molecules not found in nature as described herein. In some embodiments, the vector is preferably an IVT plasmid. In some embodiments, disclosed herein are compositions containing the nucleic acid molecules not found in nature as described herein.

[0040] In some embodiments, what is provided herein is a nucleic acid that does not exist in nature, comprising a coding nucleotide sequence encoding a protein described herein. In some embodiments, what is provided herein is a nucleic acid that does not exist in nature, comprising a coding nucleotide sequence encoding a fragment described herein. In some embodiments, what is provided herein is a nucleic acid that does not exist in nature, comprising a coding nucleotide sequence encoding a fusion protein described herein. In some embodiments, the coding nucleotide sequence is codon-optimized for expression in the cells of interest. In some embodiments, the coding nucleotide sequence is codon-optimized for expression in non-human mammalian cells. In specific embodiments, the coding nucleotide sequence is codon-optimized for expression in human cells. In some embodiments, the nucleic acid that does not exist in nature comprises a nucleotide sequence described in SEQ ID NOs: 7, 9, 11, 13, 4, or 5. The nucleic acid that does not exist in nature comprises a nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NOs: 7, 9, 11, 13, 4, or 5. In some embodiments, nucleic acids that do not exist in nature include the nucleotide sequence described in SEQ ID NO: 7.

[0041] In some embodiments, provided herein are non-naturally occurring nucleic acids encoding the proteins described herein, the proteins comprising a variant of mature gE of VZV and a human IgE signal peptide, and the nucleotide sequence encoding the IgE signal peptide comprises the nucleotide sequence described in SEQ ID NO: 24, 25, or 26.

[0042] In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding the fusion protein described herein, the fusion protein comprising a fragment of mature gE from VZV and a human IgE signal peptide, and the nucleotide sequence encoding the IgE signal peptide comprises the nucleotide sequence described in SEQ ID NO: 24, 25, or 26.

[0043] In some embodiments, provided herein are non-naturally occurring nucleic acids encoding the proteins described herein, the proteins comprising a variant of mature gE of VZV and a VZV gE signal peptide, the nucleotide sequence of the signal peptide comprising the nucleotide sequence described in SEQ ID NOs. 19, 20, 21, or 22.

[0044] In some embodiments, provided herein are non-naturally occurring nucleic acids encoding the proteins described herein, the proteins comprising a variant of mature gE of VZV and a human tPA signal peptide, and the nucleotide sequence encoding the human tPA signal peptide comprising the nucleotide sequence described in SEQ ID NO: 28. In some embodiments, provided herein are non-naturally occurring nucleic acids encoding the proteins described herein, the proteins comprising a fragment of mature gE of VZV and a human tPA signal peptide, and the nucleotide sequence encoding the human tPA signal peptide comprising a nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 28.

[0045] In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding the fusion protein described herein, the fusion protein comprising a fragment of mature gE from VZV and a human tPA signal peptide, and the nucleotide sequence encoding the human tPA signal peptide comprising the nucleotide sequence described in SEQ ID NO: 28. In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding the fusion protein described herein, the fusion protein comprising a fragment of mature gE from VZV and a human tPA signal peptide, and the nucleotide sequence encoding the human tPA signal peptide comprising a nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 28.

[0046] In some embodiments, what is provided herein is a nucleic acid that does not exist in nature, comprising the nucleotide sequence of SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, the nucleic acid that does not exist in nature comprises the nucleotide sequence of SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, the nucleic acid that does not exist in nature comprises, is essentially derived from, or includes the nucleotide sequence described in SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, nucleic acids that do not exist in nature include nucleotide sequences having 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%, or at least 99% identity to the nucleotide sequences described in SEQ ID NOs. 63, 7, 51, 60, 62, 62, 64, 9, 9, 9, 9, 9, 9, 9, or 5. In some embodiments, nucleic acids not found in nature consist of, are essentially, or include nucleotide sequences having 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%, or at least 99% identity with nucleotide sequences described in SEQ ID NOs. 63, 7, 51, 60, 62, 62, or 64. In some embodiments, nucleic acids not found in nature consist of, are essentially, or include nucleotide sequences described in SEQ ID NOs. 63. In some embodiments, nucleic acids not found in nature consist of nucleotide sequences described in SEQ ID NOs. 63.

[0047] In some embodiments, the nucleic acids described herein that do not exist in nature further include a 5' untranslated region (5'-UTR) and / or a 3' untranslated region (3'-UTR). In some embodiments, the non-naturally occurring nucleic acid described herein further comprises a 5'-UTR, the 5'-UTR comprising the nucleotide sequence described in any one of SEQ ID NOs. 29 to 38. In some embodiments, the non-naturally occurring nucleic acid described herein further comprises a 3'-UTR, the 3'-UTR comprising the nucleotide sequence described in any one of SEQ ID NOs. 39 to 46. In some embodiments, the non-naturally occurring nucleic acid described herein further comprises a 5'-UTR and a 3'-UTR, the 5'-UTR comprising the nucleotide sequence described in any one of SEQ ID NOs. 29 to 38, and the 3'-UTR comprising the nucleotide sequence described in any one of SEQ ID NOs. 39 to 46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal.

[0048] In some embodiments, the non-naturally occurring nucleic acids described herein include DNA. In some embodiments, the non-naturally occurring nucleic acids described herein are DNA. In some embodiments, the non-naturally occurring nucleic acids described herein include one or more functional nucleotide analogs. In some embodiments, the non-naturally occurring nucleic acids described herein include mRNA, in which thymine is substituted with uracil or a functional analog within the nucleic acid. In some embodiments, the non-naturally occurring nucleic acids described herein are mRNA, in which thymine is substituted with uracil or a functional analog within the nucleic acid. In some embodiments, the nucleic acid includes one or more functional nucleotide analogs selected from pseudouridine, 1-methylpsoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid consists of, is essentially, or includes the nucleotide sequence described in SEQ ID NO: 63, except that all thymine (T) is substituted with uracil (U) or N1-methylpsoiduridine. In some embodiments, the nucleic acid has all thymine (T) substituted with uracil (U) or N1-methylpsoiduridine, and the first nucleotide G is m 7In some embodiments, the nucleic acid consists of, is essentially derived from, or includes the nucleotide sequence described in SEQ ID NO: 63, except that it is substituted with GpppAmpU. In some embodiments, the nucleic acid consists of, is essentially derived from, or includes the nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 63, except that all thymine (T) is substituted with uracil (U) or N1-methylpsoiduridine. In some embodiments, the nucleic acid consists of, is essentially derived from, or includes the nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in SEQ ID NO: 63, wherein the first nucleotide G is m 7 It is substituted with GpppAmpU. In some embodiments, the nucleic acid has all thymine (T) substituted with uracil (U) or N1-methylpsoiduridine, and the first nucleotide G is m 7 The nucleic acid consists of, essentially consists of, or includes a nucleotide sequence that is 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%, or at least 99% identical to the nucleotide sequence described in Sequence ID No. 63, except that it is substituted with GpppAmpU. In some embodiments, the nucleic acid is such that all thymine (T) is substituted with N1-methylpsoiduridine, and the first nucleotide G is m 7 The nucleotide sequence described in SEQ ID NO: 63, except that it is substituted with GpppAmpU, consists of, is essentially derived from, or includes the same sequence.

[0049] In some embodiments, what is provided herein is a vector containing the nucleic acids described herein. In some embodiments, what is provided herein is a vector containing nucleic acids that do not exist in nature as described herein. In some embodiments, the vector is an IVT (in vitro transcription) plasmid.

[0050] In some embodiments, what is provided herein is a host cell containing the nucleic acid described herein. In some embodiments, what is provided herein is a host cell containing the nucleic acid not found in nature described herein. In some embodiments, what is provided herein is a host cell containing the vector described herein. In some embodiments, the host cell expresses the protein, the fragment, or the fusion protein described herein. In some embodiments, the host cell is in vitro, ex vivo, or isolated.

[0051] In some embodiments of the compositions described herein, the composition further comprises at least one lipid described herein. In some embodiments of the compositions described herein, the composition further comprises at least a first lipid described herein (e.g., a cationic lipid) and optionally a second lipid described herein (e.g., a polymer-conjugated lipid).

[0052] In some embodiments, the first lipid is a compound according to series 01, 02, 03, and 04, for example, a compound according to formula (01-I), (01-II), (02-I), (03-I), or (04-I). In some embodiments, the first lipid is a compound listed in Table 01-1, 02-1, 03-1, or 04-1. In some embodiments, the second lipid is a compound according to series 05, for example, a compound according to formula (05-I).

[0053] In some embodiments, the composition is formulated as lipid nanoparticles that encapsulate nucleic acids within a lipid shell. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the composition is a vaccine.

[0054] In some embodiments, what is provided herein is a composition (e.g., a pharmaceutical composition) comprising the protein described herein. In some embodiments, what is provided herein is a composition (e.g., a pharmaceutical composition) comprising the fragment described herein. In some embodiments, what is provided herein is a composition (e.g., a pharmaceutical composition) comprising the fusion protein described herein. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the pharmaceutical composition is a vaccine. In some embodiments, the protein, fragment, or fusion protein described herein is used to immunize a subject with VZV. In some embodiments, the nucleic acid, protein, fragment, or fusion protein described herein is used to induce an immune response in a subject (e.g., an immune response described in Section 6). In some embodiments, the immune response comprises a humoral immune response to VZV. In some embodiments, the immune response comprises a cellular response to VZV. In some embodiments, the immune response comprises an antibody specific to VZV gE (e.g., a neutralizing antibody).

[0055] In some embodiments, what is provided herein is a composition comprising the nucleic acid described herein (e.g., a pharmaceutical composition). In some embodiments, what is provided herein is a composition comprising the nucleic acid not found in nature described herein (e.g., a pharmaceutical composition). In some embodiments, what is provided herein is a composition comprising the vector described herein (e.g., a pharmaceutical composition). In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the pharmaceutical composition is a vaccine. In some embodiments, the nucleic acid, the nucleic acid not found in nature described herein, or the vector described herein is used to immunize a subject with VZV. In some embodiments, the nucleic acid, the nucleic acid not found in nature described herein, or the vector described herein is used to induce an immune response in a subject (e.g., an immune response described in Section 6). In some embodiments, the immune response comprises a humoral immune response to VZV. In some embodiments, the immune response comprises a cellular response to VZV. In some embodiments, the immune response comprises an antibody specific to VZV gE (e.g., a neutralizing antibody).

[0056] In some embodiments, what is provided herein is a pharmaceutical composition comprising a nucleic acid not found in nature as described herein and at least a first lipid. In some embodiments, the first lipid is a lipid as described herein. In some embodiments, the first lipid is a compound according to formula 01-I or formula 01-II, or a compound listed in Table 01-1, or a compound according to formula 02-I, or a compound listed in Table 02-1, or a compound according to formula 03-I, or a compound listed in Table 03-1, or a compound according to formula 04-I, or a compound listed in Table 04-1. In some embodiments, the pharmaceutical composition is as described in claim 72 and further comprises a second lipid. In some embodiments, the second lipid is a compound according to formula 05-I. In some embodiments, the pharmaceutical composition is formulated as lipid nanoparticles encapsulating the nucleic acid within a lipid shell. In some embodiments, the pharmaceutical composition is a vaccine.

[0057] In one embodiment, the foregoing provides a method for managing, preventing or treating a disease or disorder in a subject caused by or due to VZV, the method comprising administering to the subject a therapeutically effective amount of a fragment described herein, a therapeutically effective amount of a nucleic acid described herein, a therapeutically effective amount of a naturally occurring nucleic acid described herein, or a therapeutically effective amount of a pharmaceutical composition described herein.

[0058] In some embodiments of the methods described herein, the subjects are human or non-human mammals. In some embodiments, the subjects are adult humans, pediatric humans, or infant humans. In some embodiments, the subjects have a disease or disorder. In some embodiments, the subjects are at risk of or susceptible to VZV infection. In some embodiments, the subjects are elderly humans. In some embodiments, the subjects have been diagnosed as positive for VZV infection. In some embodiments, the subjects are asymptomatic.

[0059] In some embodiments of the method described herein, the method comprises administering lipid nanoparticles encapsulating nucleic acids to a subject, the lipid nanoparticles being endocytized by cells within the subject. In some embodiments, the nucleic acids are expressed by cells within the subject.

[0060] In some embodiments of the methods described herein, an immune response to VZV is induced within the subject. In some embodiments, the immune response includes the production of cytokines within lymphocytes. In some embodiments, the immune response includes an increase in the percentage of cytokine-expressing lymphocytes. In some embodiments, the lymphocytes are CD4 + T cells and / or CD8 +These are T cells. In some embodiments, the cytokines are one or more of IFN-γ, IL-2, and TNF-α. In some embodiments, cytokine production within lymphocytes is increased. In some embodiments, the immune response involves the production of antibodies that specifically bind to the viral gE protein. In some embodiments, the antibodies are neutralizing antibodies against VZV or cells infected with VZV. In some embodiments, the serum titer of the antibody is increased within the subject.

[0061] In some embodiments of the methods described herein, the antibody binds to viral particles or infected cells and labels the viral particles or infected cells so that they are destroyed by the immune system of the subject. In some embodiments, endocytosis of the viral particles to which the antibody binds is induced or enhanced. In some embodiments, antibody-dependent cell-mediated cytotoxicity (ADCC) against infected cells in the subject is induced or enhanced. In some embodiments, antibody-dependent cell phagocytosis (ADCP) against infected cells in the subject is induced or enhanced. In some embodiments, complement-dependent cytotoxicity (CDC) against infected cells in the subject is induced or enhanced.

[0062] In some embodiments of the methods described herein, the disease or disorder caused by VZV is varicella and / or herpes zoster. In some embodiments of the methods described herein, the disease or disorder caused by VZV is postherpetic neuralgia (PHN). In some embodiments of the methods described herein, the disease or disorder caused by VZV is one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcer, hepatitis, and pancreatitis.

[0063] In some embodiments, the methods provided herein are for managing, preventing or treating a disease or disorder caused by or by VZV infection in a subject, comprising administering a therapeutically effective amount of the protein described herein to the subject. In some embodiments, the methods provided herein are for managing, preventing or treating a disease or disorder caused by or by VZV infection in a subject, comprising administering a therapeutically effective amount of the fragment described herein to the subject. In some embodiments, the methods provided herein are for managing, preventing or treating a disease or disorder caused by or by VZV infection in a subject, comprising administering a therapeutically effective amount of the fusion protein described herein to the subject. In some embodiments, the methods provided herein are for managing, preventing or treating a disease or disorder caused by or by VZV infection in a subject, comprising administering a therapeutically effective amount of the nucleic acid described herein to the subject. In some embodiments, the method provided herein is for managing, preventing or treating a disease or disorder caused by or by VZV infection in a subject, comprising administering a therapeutically effective amount of a naturally occurring nucleic acid described herein to the subject. In some embodiments, the method provided herein is for managing, preventing or treating a disease or disorder caused by or by VZV infection in a subject, comprising administering a therapeutically effective amount of a vector described herein to the subject. In some embodiments, the method provided herein is for managing, preventing or treating a disease or disorder caused by or by VZV infection in a subject, comprising administering a therapeutically effective amount of a pharmaceutical composition described herein to the subject. In some embodiments, the method is for preventing a disease or disorder in a subject caused by or by VZV infection.In some embodiments, an immune response against VZV is induced in a subject. In some embodiments, the immune response includes production of cytokines in lymphocytes. In some embodiments, the immune response includes an increase in the percentage of cytokine-expressing lymphocytes. In some embodiments, the lymphocytes are CD4. + T cells and / or CD8 + T cells, and / or the cytokines are one or more of IFN-γ, IL-2, and TNF-α. In some embodiments, production of cytokines in lymphocytes increases. In some embodiments, the immune response includes production of antibodies (e.g., neutralizing antibodies) that specifically bind to VZV gE. In some embodiments, the disease or disorder caused by VZV is (a) chickenpox or herpes zoster; (b) postherpetic neuralgia (PHN); or (c) one or more of meningitis, encephalomyelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) chickenpox and herpes zoster; (b) postherpetic neuralgia (PHN); or (c) one or more of meningitis, encephalomyelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) chickenpox or herpes zoster; (b) postherpetic neuralgia (PHN); and (c) one or more of meningitis, encephalomyelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) chickenpox and herpes zoster; (b) postherpetic neuralgia (PHN); and (c) one or more of meningitis, encephalomyelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the subject is human. In some embodiments, the human is an adult human. In some embodiments, the adult is at least 40 years old. In some embodiments, the adult is at least 45 years old. In some embodiments, the adult is at least 50 years old. In some embodiments, the adult is at least 55 years old. In some embodiments, the adult is at least 60 years old. In some embodiments, the human is an elderly human.

[0064] In some embodiments, provided herein are proteins, fragments, or fusion proteins described herein for use in methods for managing, preventing, or treating diseases or disorders in a subject caused by or infection with VZV. In some embodiments, provided herein are nucleic acids, non-naturally occurring nucleic acids described herein, or vectors described herein for use in methods for managing, preventing, or treating diseases or disorders in a subject caused by or infection with VZV. In some embodiments, provided herein are pharmaceutical compositions described herein for use in methods for managing, preventing, or treating diseases or disorders in a subject caused by or infection with VZV. In some embodiments, the diseases or disorders caused by VZV are (a) varicella or herpes zoster; (b) postherpetic neuralgia (PHN); or (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) varicella and herpes zoster; (b) postherpetic neuralgia (PHN); or (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) varicella or herpes zoster; (b) postherpetic neuralgia (PHN); and (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) varicella and herpes zoster; (b) postherpetic neuralgia (PHN); and (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the subject is a human being. In some embodiments, the human being is an adult human being. In some embodiments, the adult is at least 40 years old. In some embodiments, the adult is at least 45 years old.In some embodiments, adults are at least 50 years old. In some embodiments, adults are at least 55 years old. In some embodiments, adults are at least 60 years old. In some embodiments, humans are elderly humans.

[0065] In some embodiments, what is provided herein is the use of proteins, fragments, or fusion proteins described herein for the production of a medicament for the management, prevention, or treatment of a disease or disorder in a subject caused by or due to VZV. In some embodiments, what is provided herein is the use of nucleic acids, non-naturally occurring nucleic acids, or vectors described herein for the production of a medicament for the management, prevention, or treatment of a disease or disorder in a subject caused by or due to VZV. In some embodiments, what is provided herein is the use of pharmaceutical compositions described herein for the production of a medicament for the management, prevention, or treatment of a disease or disorder in a subject caused by or due to VZV. In some embodiments, the disease or disorder caused by VZV is (a) varicella or herpes zoster; (b) postherpetic neuralgia (PHN); or (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcer, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) varicella and herpes zoster; (b) postherpetic neuralgia (PHN); or (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) varicella or herpes zoster; (b) postherpetic neuralgia (PHN); and (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the disease or disorder caused by VZV is (a) varicella and herpes zoster; (b) postherpetic neuralgia (PHN); and (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcers, hepatitis, and pancreatitis. In some embodiments, the subject is a human being. In some embodiments, the human being is an adult human being. In some embodiments, the adult is at least 40 years old. In some embodiments, the adult is at least 45 years old.In some embodiments, adults are at least 50 years old. In some embodiments, adults are at least 55 years old. In some embodiments, adults are at least 60 years old. In some embodiments, humans are elderly humans. 4. Brief explanation of the drawing [Brief explanation of the drawing]

[0066] [Figure 1] This shows the expression of candidate substances by HEK293T transfected in vitro. The suffix psd represents pseudo-U modification, and the suffix m1 represents 1-N-pseudo-U modification. [Figure 2] This shows the gE-specific IgG titer induced by the candidate substance. The suffix psd represents pseudo-U modification, and the suffix m1 represents 1-N-pseudo-U modification. [Figure 3] This shows gE peptide pool-specific T cell responses induced by candidate substances. The suffix psd represents pseudo-U modification, and the suffix m1 represents 1-N-pseudo-U modification. [Figure 4] The predicted secondary structure of full-length VZV gE is shown. A: wild type, B: mutants including substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A. [Figure 5] This shows the median fluorescence intensity of plasma-transferred cells in vitro using the candidate substance. [Figure 6] This shows the expression rate of candidate substances in plasma-transferred cells in vitro. [Figure 7] This shows the gE-specific IgG titer induced by the candidate substance. [Figure 8] This shows a gE peptide pool-specific T cell response induced by the candidate substance. [Figure 9] This diagram shows the location of the target protein within the cell. Green represents the gE protein, red represents the Golgi apparatus, and blue represents the cell nucleus. [Figure 10] This study demonstrates the in vitro analysis of RIG-I activation and IFN-β release using mRNA to suppress the inflammatory response. [Figure 11] Shows the transfection cell expression rate in vitro by the candidate substance. [Figure 12] Shows the median fluorescence intensity of the transfection cells in vitro by the candidate substance. [Figure 13A] Shows the gE-specific IgG titer after the primary immunization in naive mice. [Figure 13B] Shows the gE-specific IgG titer after the booster immunization in naive mice. [Figure 13C] Shows the gE-specific IgG titer after the primary immunization in LAV-experienced mice. [Figure 13D] Shows the gE-specific IgG titer after the booster immunization in LAV-experienced mice. [Figure 14A] Shows the CD4+ T cell response induced by the vaccine in naive mice. [Figure 14B] Shows the CD4+ T cell response induced by the vaccine in LAV-experienced mice.

Mode for Carrying Out the Invention

[0067] 5. Detailed Description Provided herein are therapeutic nucleic acid molecules useful for the prevention, management, and treatment of diseases or disorders caused by VZV or by VZV infection. Also provided herein are pharmaceutical compositions (including pharmaceutical compositions formulated as lipid nanoparticles) comprising a therapeutic nucleic acid molecule for preventing, managing, and treating diseases or disorders caused by VZV or by VZV infection, as well as related therapeutic methods and uses. Additional features of the disclosure will be apparent to those skilled in the art in view of the following detailed description of specific embodiments.

[0068] 5.1 General Techniques The techniques and procedures described or referenced herein include those commonly understood and / or used by those skilled in the art using conventional methodologies (e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual (3rd ed. 2001); Current Protocols in Molecular Biology (Ausubel et al. eds., 2003), which are widely used methodologies).

[0069] 5.2 Terminology Unless otherwise stated, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art. For the purpose of interpreting this specification, the following definitions of terms apply, and wherever appropriate, a singular term also includes its plural form, and vice versa. All patents, applications, published applications, and other publications are invoked in their entirety by reference. In the event of any conflict between a definition of a term provided herein and any document invoked by reference herein, the definitions below shall prevail.

[0070] As used herein, unless otherwise specified, the term “lipids” refers to a group of organic compounds including, but not limited to, esters of fatty acids, generally characterized by being poorly soluble in water but soluble in many nonpolar organic solvents. Lipids are generally poorly soluble in water, but there are certain categories of lipids (e.g., lipids modified with polar groups, e.g., DMG-PEG2000) that have limited water solubility and can dissolve in water under certain conditions. Known types of lipids include biomolecules such as fatty acids, waxes, sterols, fat-soluble vitamins, monoglycerides, diglycerides, triglycerides, and phospholipids. Lipids can be divided into at least three classes: (1) “simple lipids” including fats and oils as well as waxes; (2) “complex lipids” including phospholipids and glycolipids (e.g., DMPE-PEG2000); and (3) “derivative lipids” such as steroids. Furthermore, as used herein, lipids also encompass lipidoid compounds. The term "lipidoid compound," or simply "lipidoid," refers to lipid-like compounds (for example, amphiphilic compounds that have lipid-like physical properties).

[0071] The terms “lipid nanoparticles” or “LNPs” refer to particles having at least one dimension on a nanometer (nm) scale (e.g., 1 to 1,000 nm) that contain one or more lipid molecules. LNPs provided herein may further contain at least one non-lipid payload molecule (e.g., one or more nucleic acid molecules). In some embodiments, LNPs include a non-lipid payload molecule partially or completely encapsulated inside a lipid shell. In particular, in some embodiments, the payload is a negatively charged molecule (e.g., mRNA encoding a viral protein), and the lipid components of the LNP include at least one cationic lipid. While not bound by theory, it is thought that cationic lipids can interact with negatively charged payload molecules and facilitate the incorporation and / or encapsulation of the payload into the LNP during LNP formation. Other lipids that can form part of the LNPs provided herein include, but are not limited to, neutral and charged lipids, e.g., steroids, polymer-conjugated lipids, and various zwitterionic lipids. In certain embodiments, LNPs according to this disclosure include one or more lipids from the series 01, 02, 03, and 04, for example, one or more lipids from the formulas (01-I), (01-II), (02-I), (03-I), and (04-I) (and their subformulas) as described herein.

[0072] The term “cationic lipid” refers to a lipid that is positively charged at any pH value or hydrogen ion activity in its environment, or that can be positively charged in response to the pH value or hydrogen ion activity of its environment (e.g., the environment of its intended use). Thus, the term “cationic” encompasses both “permanently cationic” and “cationically cationizable.” In certain embodiments, the positive charge in a cationic lipid arises from the presence of a quaternary nitrogen atom. In certain embodiments, cationic lipids include zwitterionic lipids that exhibit a positive charge in the environment of their intended use (e.g., at physiological pH). In certain embodiments, cationic lipids are one or more lipids from the series 01, 02, 03, and 04, e.g., one or more lipids from formulas (01-I), (01-II), (02-I), (03-I), and (04-I) (and their subformulas) as described herein. The term "anionic lipid" refers to a lipid that is negatively charged at any pH value or hydrogen ion activity in its environment, or that can be negatively charged in response to the pH value or hydrogen ion activity of its environment (e.g., the environment of its intended use). An example of anionic lipids is one or more phosphate groups that have a negative charge (e.g., at physiological pH).

[0073] The term "polymer-conjugated lipid" refers to a molecule that contains both a lipid and a polymer portion. An example of a polymer-conjugated lipid is a PEG-lipid, in which the polymer portion contains polyethylene glycol.

[0074] The term "neutral lipid" encompasses any lipid molecules that exist in an uncharged or neutral amphoteric form within a selected pH value or range. In some embodiments, the selected useful pH value or range corresponds to the pH conditions in the environment of the intended use of the lipid, such as physiological pH. Examples of neutral lipids that may be used in connection with this disclosure include, but are not limited to, phosphotidylcholines such as 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC); phosphatidylethanolamines such as 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 2-((2,3-bis(oleoyloxy)propyl)dimethylammonio)ethyl hydrogen phosphate (DOCP), and sphingomyelin (SM); steroids such as ceramides and sterols, and their derivatives. The neutral lipids provided herein may be synthesized or derived (isolated or modified) from natural sources or compounds.

[0075] The term "charged lipid" encompasses any lipid molecule that exists in either a positively charged or negatively charged form at a selected pH or within a selected pH range. In some embodiments, the selected pH value or range corresponds to the pH conditions of the environment in which the lipid is intended to be used, e.g., physiological pH. Examples of neutral lipids that can be used in connection with this disclosure include, but are not limited to, phosphatidylserine, phosphatidic acid, phosphatidylglycerol, phosphatidylinositol, hemysuccinate sterols, dialkyltrimethylaluminium-propane (e.g., DOTAP, DOTMA), dialkyldimethylaminopropane, ethylphosphocholine, dimethylaminoethanecarbamoylsterol (e.g., DC-Chol), 1,2-dioleoyl-sn-glycero-3-phospho-L-serine sodium salt (DOPS-Na), 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) sodium salt (DOPG-Na), and 1,2-dioleoyl-sn-glycero-3-phosphate sodium salt (DOPA-Na). Charged lipids provided herein may be synthesized or derived (isolated or modified) from natural sources or compounds.

[0076] As used herein, unless otherwise specified, the term “alkyl” refers to a saturated linear or branched hydrocarbon chain radical consisting only of carbon and hydrogen atoms. In one embodiment, an alkyl group is, for example, a group consisting of 1 to 24 carbon atoms (C1 to C24). 24 Alkyl), 4 to 20 carbon atoms (C4~C 20 Alkyl), 6-16 carbon atoms (C6-C 16 Alkyl), 6-9 carbon atoms (C6-C9 alkyl), 1-15 carbon atoms (C1-C 15 Alkyl), 1 to 12 carbon atoms (C1 to C 12A alkyl group has 1 to 8 carbon atoms (C1-C8 alkyl) or 1 to 6 carbon atoms (C1-C6 alkyl), with the carbon atoms bonded to the rest of the molecule by single bonds. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, and 2-methylhexyl. Unless otherwise specified, alkyl groups are substituted at will.

[0077] As used herein, unless otherwise specified, the term “alkenyl” refers to a linear or branched hydrocarbon chain radical consisting only of carbon and hydrogen atoms and containing one or more carbon-carbon double bonds. The term “alkenyl” also encompasses radicals having “cis” and “trans” configurations, or alternatively, “E” and “Z” configurations, as understood by those skilled in the art. In one embodiment, the alkenyl group is, for example, 2 to 24 carbon atoms (C2 to C2). 24 Alkenyl), 4-20 carbon atoms (C4-C 20 Alkenyl), 6-16 carbon atoms (C6-C6) 16 Alkenyls), 6-9 carbon atoms (C6-C9 alkenyls), 2-15 carbon atoms (C2-C9 alkenyls), 15 Alkenyl), 2 to 12 carbon atoms (C2 to C2) 12 Alkenyl groups have 2 to 8 carbon atoms (C2-C8 alkenyls) or 2 to 6 carbon atoms (C2-C6 alkenyls), with the carbon atoms bonded to the rest of the molecule by single bonds. Examples of alkenyl groups include, but are not limited to, ethenyl, propa-1-enyl, buta-1-enyl, penta-1-enyl, and penta-1,4-dienyl. Unless otherwise specified, alkenyl groups are optionally substituted.

[0078] As used herein, unless otherwise specified, the term "alkynyl" refers to a linear or branched hydrocarbon chain radical consisting only of carbon and hydrogen atoms and containing one or more carbon-carbon triple bonds. In one embodiment, the alkynyl group is, for example, 2 to 24 carbon atoms (C2 to C2). 24 Alkynyl), 4 to 20 carbon atoms (C4~C 20 Alkynyl), 6-16 carbon atoms (C6-C6) 16 Alkynyl), 6-9 carbon atoms (C6-C9 alkynyl), 2-15 carbon atoms (C2-C 15 Alkynyl), 2 to 12 carbon atoms (C2 to C2) 12 Alkynyl groups have 2 to 8 carbon atoms (C2-C8 alkynyl) or 2 to 6 carbon atoms (C2-C6 alkynyl), with the carbon atoms bonded to the rest of the molecule by single bonds. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, and pentynyl. Unless otherwise specified, alkynyl groups are optionally substituted.

[0079] As used herein, unless otherwise specified, the terms “alkylene” or “alkylene chain” refer to a saturated linear or branched divalent hydrocarbon chain in which the remainder of the molecule is linked to radical groups consisting only of carbon and hydrogen. In one embodiment, the alkylene is, for example, 1 to 24 carbon atoms (C1 to C24). 24 Alkylene) has 1 to 15 carbon atoms (C1 to C 15 Alkylene), 1 to 12 carbon atoms (C1 to C 12Alkylenes have 1 to 8 carbon atoms (C1-C8 alkylenes), 1 to 6 carbon atoms (C1-C6 alkylenes), 2 to 4 carbon atoms (C2-C4 alkylenes), and 1 to 2 carbon atoms (C1-C2 alkylenes). Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, and n-butylene. Alkylene chains are bonded to the rest of the molecule via single bonds and to radical groups via single bonds. Bonding sites of alkylene chains to the rest of the molecule and to radical groups can be via one carbon or any two carbons in the chain. Unless otherwise specified, alkylene chains are optionally substituted.

[0080] As used herein, unless otherwise specified, the term “alkenylene” refers to a linear or branched divalent hydrocarbon chain containing one or more carbon-carbon double bonds, with the remainder of the molecule linked to a radical group consisting solely of carbon and hydrogen. In one embodiment, the alkenylene is, for example, 2 to 24 carbon atoms (C2 to C2). 24 Alkenylenes), 2 to 15 carbon atoms (C2 to C2) 15 Alkenylenes), 2 to 12 carbon atoms (C2 to C2) 12 Alkenylenes have 2 to 8 carbon atoms (C2-C8 alkenylenes), 2 to 6 carbon atoms (C2-C6 alkenylenes), or 2 to 4 carbon atoms (C2-C4 alkenylenes). Examples of alkenylenes include, but are not limited to, etenylene, propenylene, and n-butenylene. Alkenylenes are bonded to the rest of the molecule via single or double bonds and to radical groups via single or double bonds. Bonding points of alkenylenes to the rest of the molecule and to radical groups can be via one carbon or any two carbons in the chain. Unless otherwise specified, alkenylenes are optionally substituted.

[0081] As used herein, unless otherwise specified, the term “cycloalkyl” refers to a saturated, non-aromatic monocyclic or polycyclic hydrocarbon radical consisting only of carbon and hydrogen atoms. Cycloalkyl groups may include fused or bridging ring systems. In one embodiment, a cycloalkyl group may, for example, consist of 3 to 15 ring carbon atoms (C3 to C3). 15 Cycloalkyl), 3-10 ring carbon atoms (C3-C 10 A cycloalkyl group has 3 to 8 ring carbon atoms (C3-C8 cycloalkyl). The cycloalkyl group is attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkyl radicals include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic cycloalkyl radicals include, but are not limited to, adamantyl, norbornyl, dekalinyl, and 7,7-dimethyl-bicyclo[2.2.1]heptanyl. Unless otherwise specified, the cycloalkyl group is optionally substituted.

[0082] As used herein, unless otherwise specified, the term "cycloalkylene" refers to a divalent cycloalkyl group. Unless otherwise specified, the cycloalkylene group is optionally substituted.

[0083] As used herein, unless otherwise specified, the term “cycloalkenyl” refers to a non-aromatic monocyclic or polycyclic hydrocarbon radical consisting only of carbon and hydrogen atoms and containing one or more carbon-carbon double bonds. Cycloalkenyls may include fused or bridging ring systems. In one embodiment, a cycloalkenyl is, for example, a ring containing 3 to 15 carbon atoms (C3 to C3). 15 Cycloalkenyl), 3-10 ring carbon atoms (C3-C 10A monocyclic cycloalkenyl radical has 3 to 8 ring carbon atoms (C3-C8 cycloalkenyl). The cycloalkenyl is attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkenyl radicals include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Unless otherwise specified, the cycloalkenyl group is optionally substituted.

[0084] As used herein, unless otherwise specified, the term "cycloalkenylene" refers to a divalent cycloalkenyl group. Unless otherwise specified, the cycloalkenylene group is optionally substituted.

[0085] As used herein, unless otherwise specified, the term “heterocyclyl” refers to a non-aromatic radical monocyclic or polycyclic moiety containing one or more heteroatoms (e.g., one, one or two, one to three, or one to four) independently selected from nitrogen, oxygen, phosphorus, and sulfur. Heterocyclyls may be bonded to the main structure with any heteroatom or carbon atom. Heterocyclyl groups can be monocyclic, bicyclic, tricyclic, tetracyclic, or other polycyclic ring systems, and polycyclic ring systems can be fused, bridging, or spirocyclic. Heterocyclyl polycyclic ring systems may contain one or more heteroatoms in one or more rings. Heterocyclyl groups may be saturated or partially unsaturated. Saturated heterocycloalkyl groups may be referred to as “heterocycloalkyl.” A partially unsaturated heterocycloalkyl group may be called a "heterocycloalkenyl" if the heterocyclyl contains at least one double bond, or a "heterocycloalkynyl" if the heterocyclyl contains at least one triple bond. In one embodiment, the heterocyclyl has, for example, 3 to 18 ring atoms (3 to 18-membered heterocyclyl), 4 to 18 ring atoms (4 to 18-membered heterocyclyl), 5 to 18 ring atoms (5 to 18-membered heterocyclyl), 4 to 8 ring atoms (4 to 8-membered heterocyclyl), or 5 to 8 ring atoms (5 to 8-membered heterocyclyl). Wherever it appears herein, numerical ranges such as "3 to 18" refer to each integer within a given range. For example, "3 to 18 ring atoms" means that a heterocyclyl group may consist of 3, 4, 5, 6, 7, 8, 9, 10 ring atoms, and may contain up to 18 ring atoms. Examples of heterocyclyl groups include, but are not limited to, imidazolyl, imidazolidinyl, oxazolyl, oxazolidinyl, thiazolyl, thiazolidinyl, pyrazolidinyl, pyrazolyl, isoxazolidinyl, isoxazolyl, isothiazolyl, isothiazolidinyl, isothiazolyl, morpholinyl, pyrrolyl, pyrrolidinyl, furyl, tetrahydrofuryl, thiophenyl, pyridinyl, piperidinyl, quinolyl, and isoquinolyl. Unless otherwise specified, heterocyclyl groups are substituted at the discretion of the user.

[0086] As used herein, unless otherwise specified, the term "heterocycle" is a divalent heterocyclyl group. Unless otherwise specified, the heterocycle group is optionally substituted.

[0087] As used herein, unless otherwise specified, the term "aryl" refers to a monocyclic aromatic group and / or a polycyclic monovalent aromatic group containing at least one aromatic hydrocarbon ring. In certain embodiments, aryl has 6 to 18 ring carbon atoms (C6-C 18 aryl), 6 to 14 ring carbon atoms (C6-C 14 aryl), or 6 to 10 ring carbon atoms (C6-C 10 aryl). Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. The term "aryl" also refers to bicyclic, tricyclic, or other polycyclic hydrocarbon rings in which at least one of the rings is aromatic and the others can be saturated, partially unsaturated, or aromatic, such as dihydronaphthyl, indenyl, indanyl, or tetrahydronaphthyl (tetralinyl). Unless otherwise specified, the aryl group is optionally substituted.

[0088] As used herein, unless otherwise specified, the term "arylene" is a divalent aryl group. Unless otherwise specified, the arylene group is optionally substituted.

[0089] As used herein, unless otherwise specified, the term “heteroaryl” means a monocyclic and / or polycyclic aromatic group containing at least one aromatic ring, the at least one aromatic ring independently containing one or more heteroatoms (e.g., one, one or two, one to three, or one to four) selected from O, S, and N. A heteroaryl can be bonded to the main structure by any heteroatom or carbon atom. In certain embodiments, a heteroaryl has 5 to 20, 5 to 15, or 5 to 10 ring atoms. The term “heteroaryl” also means a bicyclic, tricyclic, or other polycyclic ring, the at least one of which is aromatic and the others may be saturated, partially unsaturated, or aromatic, the at least one aromatic ring independently containing one or more heteroatoms selected from O, S, and N. Examples of monocyclic heteroaryl groups include, but are not limited to, pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridadinyl, and triazinyl. Examples of bicyclic heteroaryl groups include, but are not limited to, indolyl, benzothiazolyl, benzoxazolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolidinyl, benzofuranyl, isobenzofuranyl, chromonyl, coumalinyl, sinnolinyl, quinoxalinyl, indazolyl, purinyl, pyrrolopyridinyl, flupyridinyl, thienopyridinyl, dihydroisoindolyl, and tetrahydroquinolinyl. Examples of tricyclic heteroaryl groups include, but are not limited to, carbazolyl, benzindolyl, phenanthrolinyl, acridinyl, phenanthridine, and xanthenyl. Unless otherwise specified, heteroaryl groups are optionally substituted.

[0090] As used herein, unless otherwise specified, the term "heteroarylene" refers to a divalent heteroaryl group. Unless otherwise specified, the heteroarylene group is optionally substituted.

[0091] Where a group described herein is said to be “substituted,” such group may be substituted with any suitable substituent(s)(s)(s). Exemplary examples of substituents include those found in the exemplary compounds and embodiments provided herein, as well as halogen atoms such as F, CI, Br, or I; cyano; oxo(=O); hydroxyl(-OH); alkyl; alkenyl; alkynyl; cycloalkyl; aryl; -(C=O)OR'; -O(C=O)R'; -C(=O)R'; -OR'; -S(O) x R';-S-SR';-C(=O)SR';-SC(=O)R';-NR'R';-NR'C(=O)R';-C(=O)NR'R';-NR'C(=O)NR'R';-OC(=O)NR'R';-NR'C(=O)OR';-NR'S(O) x NR'R';-NR'S(O) x R'; and -S(O) x NR'R'(wherein R' is independent of H and C1~C in each occurrence) 15 Examples include, but are not limited to, alkyl or cycloalkyl groups (where x is 0, 1, or 2). In some embodiments, the substituents are C1-C 12 In other embodiments, the substituent is an alkyl group. In other embodiments, the substituent is a cycloalkyl group. In other embodiments, the substituent is a halo group such as a fluoro group. In other embodiments, the substituent is an oxo group. In other embodiments, the substituent is a hydroxyl group. In other embodiments, the substituent is an alkoxy group (-OR'). In other embodiments, the substituent is a carboxyl group. In other embodiments, the substituent is an amino group (-NR'R').

[0092] As used herein, and unless otherwise specified, the terms “optional” or “optionally” (e.g., optionally substituted) mean that the events of the situation described thereafter may or may not occur, and that the description includes both instances in which such events or situations occur and instances in which they do not. For example, “optionally substituted alkyl” means that the alkyl radical may or may not be substituted, and the description includes both substituted and unsubstituted alkyl radicals.

[0093] As used herein, unless otherwise specified, the term “prodrug” of a biologically active compound refers to a compound that can be converted to a biologically active compound under physiological conditions or by dissolution. In one embodiment, the term “prodrug” refers to a metabolic precursor of a pharmaceutically acceptable biologically active compound. A prodrug may be inactive when administered to a target that requires it, but is converted to a biologically active compound in vivo. Prodrugs are typically rapidly converted in vivo, for example, by hydrolysis in the blood, to produce the original biologically active compound. Prodrug compounds often offer advantages in solubility, histocompatibility, or delayed release in mammalian organisms (see Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). Discussions of prodrugs are provided in Higuchi, T., et al., ACS Symposium Series, Vol. 14, and Bioreversible Carriers in Drug Design, Ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

[0094] In one embodiment, the term “prodrug” also means that such a prodrug includes any covalent carrier that releases the active compound in vivo when administered to a mammalian subject. A prodrug of a compound can be prepared by modifying a functional group present in the compound in such a manner that the modification is cleaved either by conventional means or in vivo to become the original compound. A prodrug includes a compound to which a hydroxyl group, an amino group, or a mercapto group is bonded, which is cleaved when the prodrug of a compound is administered to a mammalian subject to form a free hydroxyl group, a free amino group, or a free mercapto group, respectively.

[0095] Examples of prodrugs include, but are not limited to, alcohol acetates, formates, and benzoate derivatives, or amide derivatives of amine functional groups, of the compounds provided herein.

[0096] As used herein, unless otherwise specified, the term “pharmaceutically acceptable salt” includes both acid and base addition salts.

[0097] Examples of pharmaceutically acceptable acid addition salts include, but are not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonate, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, and glutamine. Examples of organic acids include acids, glutaric acid, 2-oxo-glutaric acid, glycerophosphate, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucinic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid.

[0098] Examples of pharmaceutically acceptable base additions include, but are not limited to, salts prepared by adding an inorganic or organic base to a free acid compound. Examples of salts derived from inorganic bases include, but are not limited to, salts of sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. In one embodiment, the inorganic salts are salts of ammonium, sodium, potassium, calcium, and magnesium. Examples of salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and salts of basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydravamin, choline, betaine, benetamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, and polyamine resins. In one embodiment, the organic base is isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0099] The compounds provided herein may contain one or more chiral centers and thus may give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined from the viewpoint of absolute stereochemistry as (R)- or (S)-, or for amino acids as (D)- or (L)-. Unless otherwise specified, the compounds provided herein include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or they can be resolved using conventional techniques, such as chromatography and fractional crystallization. Conventional techniques for preparing / isolating individual enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography (HPLC). Where a compound described herein contains an olefin double bond or other geometrically asymmetric center, unless otherwise specified, the compound is intended to include both E and Z geometric isomers. Similarly, all tautomer forms are also intended to be included.

[0100] As used herein, and unless otherwise specified, the term “isomer” refers to different compounds having the same molecular formula. “Stereoisomers” are isomers that differ only in the way their atoms are arranged in space. “Atropisomers” are stereoisomers due to binding rotations around a single bond. “Enantiomers” are a pair of stereoisomers that are mirror images of each other and cannot be superimposed. A mixture of any proportion of a pair of enantiomers may be known as a “racemic” mixture. “Diastereoisomers” are stereoisomers that have at least two chiral atoms but are not mirror images of each other.

[0101] "Stereoisomers" may also include E and Z isomers, or mixtures thereof, as well as cis and trans isomers, or mixtures thereof. In certain embodiments, the compounds described herein are isolated as either the E or Z isomer. In other embodiments, the compounds described herein are mixtures of the E and Z isomers.

[0102] A "tautomer" refers to an isomer of a compound that exists in equilibrium with it. The concentration of the isomer depends on the environment in which the compound is found, and may differ depending on whether the compound is a solid or in an organic solution or aqueous solution.

[0103] It should also be noted that the compounds described herein may contain one or more atomic isotopes in unnatural proportions. For example, a compound may contain tritium ( 3 H), Iodine-125 ( 125 I), Sulfur 35 ( 35 S), or carbon-14 ( 14 Even if radioactively labeled with radioactive isotopes such as C, or deuterium ( 2 H), carbon-13 ( 13 C), or nitrogen 15 ( 15Isotopes such as N) may be enriched. As used herein, “isotopolog” refers to an isotope-enriched compound. The term “isotopically enriched” refers to an atom having an isotope composition other than the natural isotope composition of that atom. “Isotope enriched” may also refer to a compound containing at least one atom having an isotope composition other than the natural isotope composition of that atom. The term “isotopically enriched” refers to the amount of each isotope present for a given atom. Radiolabeled and isotope-enriched compounds are useful as therapeutic agents (e.g., cancer treatments), research reagents (e.g., binding assay reagents), and diagnostic agents (e.g., in vivo contrast agents). All isotope variations of the compounds described herein, whether radioactive or not, are intended to be included within the scope of the embodiments provided herein. In some embodiments, isotope molecular species of the compounds described herein are provided, for example, isotope molecular species enriched with deuterium, carbon-13, and / or nitrogen-15. As used herein, “deuterated” means that at least one hydrogen (H) is converted into deuterium (D or 2 It is replaced by (represented by H), meaning that deuterium is concentrated at at least one position in the compound.

[0104] In cases where there is a discrepancy between the illustrated structure and its name, please note that the illustrated structure should be followed.

[0105] As used herein and unless otherwise specified, the term “pharmaceutically acceptable carrier, diluent or excipient” includes, but is not limited to, any adjuvants, carriers, excipients, lubricants, sweeteners, diluents, preservatives, colorants, flavor enhancers, surfactants, humectants, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers that are approved by the U.S. Food and Drug Administration (FDA) as acceptable for use in human or animal husbandry.

[0106] The term “composition” is intended to encompass products containing, optionally, a specified amount of a specified component (e.g., the mRNA molecule provided herein).

[0107] As used herein interchangeably, the terms “polynucleotide” and “nucleic acid” refer to polymers of nucleotides of any length, including, for example, DNA and RNA. A nucleotide may be a deoxyribonucleotide, ribonucleotide, modified nucleotide or base, and / or analogues thereof, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthetic reaction. Polynucleotides may include modified nucleotides (e.g., methylated nucleotides and their analogues). Nucleic acids may be single-stranded or double-stranded. As used herein, and unless otherwise specified, “nucleic acid” also includes nucleic acid mimics such as loc nucleic acid (LNA), peptide nucleic acid (PNA), and morpholino. As used herein, “oligonucleotide” refers to a short synthetic polynucleotide, which is generally less than approximately 200 nucleotides in length, though not necessarily so. The terms “oligonucleotide” and “polynucleotide” are not mutually exclusive. The above description of polynucleotides is equally and fully applicable to oligonucleotides. Unless otherwise specified, the left-hand end of any single-stranded polynucleotide sequence disclosed herein is the 5' end, and the left-hand direction of a double-stranded polynucleotide sequence is referred to as the 5' direction. The direction of addition from 5' to 3' of a nascent RNA transcript is referred to as the transcription direction, the sequence region on the DNA strand having the same sequence as the RNA transcript, located 5' to the 5' end of the RNA transcript, is referred to as the "upstream sequence," and the sequence region on the DNA strand having the same sequence as the RNA transcript, located 3' to the 3' end of the RNA transcript, is referred to as the "downstream sequence."

[0108] As used herein, the term “not naturally occurring” is intended to mean, when used in reference to a nucleic acid molecule described herein, that nucleic acid molecule is not found in nature. A nucleic acid not naturally occurring that codes for a viral peptide or protein contains at least one genetic alteration or chemical modification not typically found in naturally occurring viral strains, including wild-type viral strains. Genetic alterations include, for example, modifications introducing an expressible nucleic acid sequence coding for a peptide or polypeptide of a different species than the virus, other nucleic acid additions, nucleic acid deletions, nucleic acid substitutions, and / or other functional disruptions of the viral genetic material. Such modifications include, for example, modifications in the coding region and functional fragments of a polypeptide that is heterogeneous, homogeneous, or both heterogeneous and homogeneous to the viral species. Additional modifications include, for example, modifications in non-coding regulatory regions that alter the expression of a gene or operon. Additional modifications also include, for example, the incorporation of a nucleic acid sequence into a vector such as a plasmid or artificial chromosome. Chemical modifications include, for example, one or more functional nucleotide analogs described herein.

[0109] "Isolated nucleic acids" are nucleic acids, such as RNA, DNA, or mixed nucleic acids, substantially separated from other genomic DNA sequences and proteins or complexes, such as ribosomes and polymerases, that naturally accompany native sequences. "Isolated" nucleic acid molecules are nucleic acid molecules that have been separated from other nucleic acid molecules present in the native source of the nucleic acid molecule. Furthermore, "isolated" nucleic acid molecules, such as mRNA molecules, may substantially contain no other cellular material or culture medium if produced by recombinant techniques, or substantially contain no chemical precursors or other chemicals if chemically synthesized. In specific embodiments, one or more nucleic acid molecules encoding the antigens described herein are isolated or purified. This term encompasses nucleic acid sequences taken from their naturally occurring environment, recombinant or cloned DNA or RNA isolates, and chemically synthesized analogs or analogs biologically synthesized by heterologous systems. A substantially pure molecule may include the isolated form of the molecule.

[0110] When used in reference to nucleic acid molecules, the term “coding nucleic acid” or its grammatical equivalent encompasses (a) nucleic acid molecules in their natural state, or, when manipulated by methods well known to those skilled in the art, nucleic acid molecules that can be transcribed to produce mRNA, which is then translated into peptides and / or polypeptides, and (b) the mRNA molecule itself. The antisense strand is the complement of such a nucleic acid molecule from which the coding sequence can be inferred. The term “coding region” refers to the portion of the coding nucleic acid sequence that is translated into peptides or polypeptides. The term “untranslated region” or “UTR” refers to the portion of the coding nucleic acid that is not translated into peptides or polypeptides. Depending on the orientation of the UTR relative to the coding region of the nucleic acid molecule, the UTR is referred to as the 5'-UTR if it is located at the 5' end of the coding region, and the UTR is referred to as the 3'-UTR if it is located at the 3' end of the coding region.

[0111] As used herein, the term "mRNA" refers to a message RNA molecule comprising one or more open reading frames (ORFs) that can be translated by a cell or organism containing mRNA to produce one or more peptide or protein products. The region containing one or more ORFs is referred to as the coding region of the mRNA molecule. In certain embodiments, the mRNA molecule further comprises one or more untranslated regions (UTRs).

[0112] In certain embodiments, the mRNA is a monocistronic mRNA containing only one ORF. In certain embodiments, the monocistronic mRNA encodes a peptide or protein containing at least one epitope of a selected antigen (e.g., a pathogenic antigen or a tumor-associated antigen). In other embodiments, the mRNA is a multicistronic mRNA containing two or more ORFs. In certain embodiments, the multicistronic mRNA encodes two or more peptides or proteins, which may be the same or different from each other. In certain embodiments, each peptide or protein encoded by the multicistronic mRNA contains at least one epitope of a selected antigen. In certain embodiments, each different peptide or protein encoded by the multicistronic mRNA contains at least one epitope of a different antigen. In any of the embodiments described herein, the at least one epitope may be at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten epitopes of the antigen.

[0113] The term "nucleic acid bases" encompasses purines and pyrimidines, including the natural compounds adenine, thymine, guanine, cytosine, uracil, inosine, and their natural or synthetic analogs or derivatives.

[0114] As used herein, the term “functional nucleotide analog” means a modified version of a standard nucleotide A, G, C, U, or T that (a) retains the base-pairing properties of the corresponding standard nucleotide, and (b) contains at least one chemical modification of the corresponding native nucleotide to (i) a nucleic acid base, (ii) a sugar group, (iii) a phosphate group, or (iv) any combination of (i) to (iii). As used herein, base pairing includes not only standard Watson-Crick type adenine-thymine, adenine-uracil, or guanine-cytosine base pairs, but also base pairs formed between a standard nucleotide and a functional nucleotide analog or between a pair of functional nucleotide analogs, where the arrangement of hydrogen bond donors and hydrogen bond acceptors allows for hydrogen bonding between a modified nucleic acid base and a standard nucleic acid base or between two complementary nucleic acid base structures. For example, a functional analog of guanosine (G) retains the ability to base pair with cytosine (C) or a functional analog of cytosine. An example of such non-standard base pairing is the base pairing between the modified nucleotide inosine and adenine, cytosine, or uracil. As described herein, functional nucleotide analogs may or may not be naturally occurring. Therefore, nucleic acid molecules containing functional nucleotide analogs may have at least one modified nucleic acid base, sugar group, and / or nucleoside linkage. Exemplary chemical modifications to nucleic acid bases, sugar groups, or nucleoside linkages of nucleic acid molecules are provided herein.

[0115] As used herein, the terms “translation-enhancer element,” “TEE,” and “translation enhancer” refer to regions within a nucleic acid molecule that function to facilitate translation from a coding sequence of a nucleic acid to a protein or peptide product, for example, via cap-dependent or cap-independent translation. TEEs are typically located within the UTR region of a nucleic acid molecule (e.g., mRNA) and enhance the translation level of coding sequences located upstream or downstream. For example, a TEE in the 5'-UTR of a nucleic acid molecule may be located between the promoter and the start codon of the nucleic acid molecule. Various TEE sequences are known in the art (Wellensiek et al. Genome-wide profiling of human cap-independent translation-enhancing elements, Nature Methods, 2013 Aug;10(8):747-750; Chappell et al. PNAS June 29, 2004 101(26)9590-9594). Some TEEs are known to be conserved across multiple species (Panek et al. Nucleic Acids Research, Volume 41, Issue 16, 1 September 2013, Pages 7625-7634).

[0116] As used herein, the term “stem-loop sequence” refers to a single-stranded polynucleotide sequence having at least two regions that are complementary or substantially complementary to each other when read in opposite directions, and are therefore capable of base-pairing with each other to form at least one double helix and one unpaired loop. The resulting structure is known as a stem-loop structure, hairpin, or hairpin loop, and is a secondary structure found in many RNA molecules.

[0117] As used herein, the term “peptide” refers to a polymer containing 2 to 50 amino acid residues linked by one or more covalent peptide bonds. This term applies to naturally occurring amino acid polymers and amino acid polymers in which one or more amino acid residues are amino acids that do not exist naturally (e.g., amino acid analogs or unnatural amino acids).

[0118] The terms “polypeptide” and “protein” are used interchangeably herein to refer to polymers of more than 50 amino acid residues linked by covalent peptide bonds. That is, descriptions of polypeptides are equally applicable to descriptions of proteins, and vice versa. The term applies to naturally occurring amino acid polymers, as well as amino acid polymers in which one or more amino acid residues are non-natural amino acids (e.g., amino acid analogs). As used herein, the term encompasses amino acid chains of any length, including full-length proteins (e.g., antigens).

[0119] In the context of peptides or polypeptides, as used herein, the term “derivative” refers to a peptide or polypeptide comprising the amino acid sequence of a viral peptide or protein, or a fragment of a viral peptide or protein, that has been altered by the introduction of substitution, deletion, or addition of amino acid residues. As used herein, the term “derivative” also refers to a viral peptide or protein, or a fragment of a viral peptide or protein, that has been chemically modified (for example, by covalently bonding any type of molecule to the polypeptide). For example, but not limited to, a viral peptide or protein, or a fragment of a viral peptide or protein, may be chemically modified by, for example, glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, chemical cleavage, formulation, metabolic synthesis of tunicamycin, linkage with cellular ligands or other proteins, etc. Derivatives are modified in a way that differs from naturally occurring or starting peptides or polypeptides, in terms of the type or position of the molecule they are bonded to. Derivatives further include deletions of one or more chemical groups that are naturally occurring on the viral peptide or protein. Furthermore, derivatives of viral peptides or proteins, or fragments of viral peptides or proteins, may contain one or more non-classical amino acids. In specific embodiments, the derivative is a functional derivative of the derived natural or unmodified peptide or polypeptide.

[0120] The term "functional derivative" refers to a derivative that retains one or more functions or activities of the naturally occurring or starting peptide or polypeptide from which it originates. For example, a functional derivative of the VZV S protein may retain the ability to bind to one or more of its receptors on a host cell. For example, a functional derivative of the VZV N protein may retain the ability to bind to RNA or packaged viral genomes.

[0121] The term "identity" refers to the relationship between two or more polypeptide molecules or two or more nucleic acid molecules determined by sequence alignment and comparison. The "amino acid sequence identity percentage (%)" relative to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in the reference polypeptide sequence after the sequences have been aligned, gaps introduced, and, if necessary, the maximum sequence identity percentage has been achieved, with no conservative substitutions being considered as part of the sequence identity. Alignment for the purpose of determining the amino acid sequence identity percentage can be achieved in various ways within the scope of the skill of a person skilled in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN (DNAStar, Inc.) software. A person skilled in the art can determine appropriate parameters for sequence alignment, including any algorithms required to achieve the maximum alignment over the full length of the sequences being compared.

[0122] "Modification" of an amino acid residue / position refers to a change in the primary amino acid sequence compared to the starting amino acid sequence, and the change results from a sequence alteration that includes amino acid residues / positions. Typical modifications include the substitution of a residue with another amino acid (e.g., conserved or non-conservative substitution), the insertion of one or more (typically five, four, or fewer than three) amino acids adjacent to a residue / position, and / or the deletion of a residue / position.

[0123] In the context of peptides or polypeptides, as used herein, the term “fragment” refers to a peptide or polypeptide containing an amino acid sequence shorter than the full-length amino acid sequence. Such fragments may arise, for example, from truncation at the amino terminus, truncation at the carboxy terminus, and / or internal deletion of residues(s) from the amino acid sequence. Fragments may also arise, for example, from alternative RNA splicing or in vivo protease activity. In certain embodiments, the fragment may consist of at least five consecutive amino acid residues, at least ten consecutive amino acid residues, at least fifteen consecutive amino acid residues, at least twenty consecutive amino acid residues, at least twenty-five consecutive amino acid residues, at least thirty consecutive amino acid residues, at least forty consecutive amino acid residues, at least fifty consecutive amino acid residues, at least sixty consecutive amino acid residues, at least seventy consecutive amino acid residues, at least eighty consecutive amino acid residues, at least ninety consecutive amino acid residues, or at least one hundred consecutive amino acid residues from the amino acid sequence of the polypeptide. This refers to a polypeptide comprising an amino acid sequence of at least 125 consecutive amino acid residues, at least 150 consecutive amino acid residues, at least 175 consecutive amino acid residues, at least 200 consecutive amino acid residues, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900, or at least 950 consecutive amino acid residues. In specific embodiments, a polypeptide fragment retains at least one, at least two, at least three, or more functions of the polypeptide.

[0124] When used herein in the context of peptides or polypeptides (e.g., proteins), the term “immunogenic fragment” refers to a fragment of a peptide or polypeptide that retains the ability of the peptide or polypeptide to induce an immune response (including innate and / or adaptive immune responses) upon contact with the mammalian immune system. In some embodiments, the immunogenic fragment of a peptide or polypeptide may be an epitope.

[0125] The term "antigen" refers to a substance that can be recognized by the immune system of a target (including the adaptive immune system) and that can trigger an immune response (including an antigen-specific immune response) after the target comes into contact with the antigen. In certain embodiments, an antigen is a protein associated with a pathogen or a pathological cell, such as a cell infected with a tumor cell (e.g., a tumor-associated antigen (TAA)).

[0126] An "epitope" is a site on the surface of an antigen molecule to which a single antibody molecule binds, such as a localized region on the surface of an antigen that can bind to one or more antigen-binding regions of an antibody and has antigenic or immunogenic activity that can induce an immune response in animals such as mammals (e.g., humans). An immunogenic epitope is a portion of a polypeptide that induces an antibody response in animals. An antigenic epitope is a portion of a polypeptide to which an antibody binds, determined by any method known in the art, including immunoassays, for example. Antigenic epitopes do not necessarily have to be immunogenic. Epitopes often consist of a chemically active surface population of molecules such as amino acids or sugar side chains and have specific three-dimensional structural properties as well as specific charge properties. Antibody epitopes may be linear epitopes or conformational epitopes. Linear epitopes are formed by a continuous sequence of amino acids in a protein. Structural epitopes are formed by amino acids that are discontinuous in the protein sequence but come together when the protein folds into its three-dimensional structure. Inducible epitopes are formed when the protein's three-dimensional structure is altered, such as after activation or binding to another protein or ligand. In certain embodiments, epitopes are three-dimensional surface features of a polypeptide. In other embodiments, epitopes are linear features of a polypeptide. Generally, antigens have several or many different epitopes and can react with many different antibodies.

[0127] The term "heterogeneous" refers to entities not found in nature that are associated with naturally occurring VZV (e.g., those encoded and / or expressed by their genome). The term "homonymous" refers to entities found in nature that are associated with naturally occurring VZV (e.g., those encoded and / or expressed by their genome).

[0128] As used herein, the term “gene vaccine” refers to a therapeutic or prophylactic composition comprising at least one nucleic acid molecule encoding an antigen associated with a target disease (e.g., an infectious disease or a neoplastic disease). Administration of the vaccine to a subject (“vaccination”) enables the production of the encoded peptide or protein, thereby inducing an immune response in the subject to the target disease. In certain embodiments, the immune response includes adaptive immune responses such as the production of antibodies against the encoded antigen and / or the activation and proliferation of immune cells that can specifically eliminate diseased cells expressing the antigen. In certain embodiments, the immune response further includes innate immune responses. According to this disclosure, the vaccine may be administered to a subject either before or after the onset of clinical symptoms of the target disease. In some embodiments, vaccination of a healthy or asymptomatic subject makes the vaccinated subject immune to or reduces susceptibility to the development of the target disease. In some embodiments, vaccination of a subject exhibiting symptoms of the disease improves the disease condition of the vaccinated subject or treats the disease.

[0129] The term "vector" refers to a substance used to contain or to contain a nucleic acid sequence (e.g., a nucleic acid sequence encoding a viral peptide or protein as described herein) for the purpose of introducing the nucleic acid sequence into a host cell or serving as a transcription template for an in vitro transcription reaction in a cell-free system for mRNA production. Applicable vectors for use include, for example, expression vectors, plasmids, phage vectors, viral vectors, episomes, and artificial chromosomes, which may contain selectable sequences or markers that can be stably incorporated into the chromosomes of a host cell. Furthermore, a vector may contain one or more selectable marker genes and appropriate transcription or translational regulatory sequences. Selectable marker genes that may be included may, for example, confer resistance to antibiotics or toxins, compensate for nutritional requirement deficiencies, or supplement essential nutrients not present in the culture medium. Examples of transcription or translational regulatory sequences include constitutive and inductive promoters, transcriptional enhancers, transcriptional terminators, etc., which are well known in the art. When two or more nucleic acid molecules are co-transcribed or co-translated (e.g., two or more nucleic acid molecules encoding different viral peptides or proteins), both nucleic acid molecules may be inserted, for example, into a single expression vector or separate expression vectors. For single-vector transcription and / or translation, the coding nucleic acid may be operationally ligated to one common transcriptional or translational regulatory sequence, or to different transcriptional or translational regulatory sequences (e.g., one inductive promoter and one constitutive promoter). The introduction of the nucleic acid molecule into host cells can be confirmed using methods well known in the art. Such methods include, for example, nucleic acid analysis such as Northern blotting or polymerase chain reaction (PCR) amplification of mRNA, immunoblotting for gene product expression, or other suitable analytical methods for testing the expression of the introduced nucleic acid sequence or the corresponding gene product. It will be understood by those skilled in the art that the nucleic acid molecule is expressed in an amount sufficient to produce the desired product (e.g., the mRNA transcript of the nucleic acid described herein), and further, that the expression level can be optimized to obtain sufficient expression using methods well known in the art.

[0130] The terms “innate immune response” and “innate immunity” are recognized in the art and refer to the nonspecific defense mechanisms initiated by the body’s immune system upon recognition of pathogen-associated molecular patterns, which involve various forms of cellular activity, including cytokine production and cell death via various pathways. As used herein, an innate immune response includes, but is not limited to, increased production of inflammatory cytokines (e.g., type I interferon or IL-10 production), activation of the NFκB pathway, increased proliferation, maturation, differentiation and / or survival of immune cells, and, if applicable, induction of cellular apoptosis. Activation of innate immunity can be detected using methods known in the art, such as the measurement of (NF)-κB activation.

[0131] The terms “adaptive immune response” and “adaptive immunity” are recognized in the art and refer to antigen-specific defense mechanisms initiated by the body’s immune system upon recognition of a particular antigen, including both humoral and cell-mediated responses. As used herein, an adaptive immune response includes cellular responses induced and / or amplified by vaccine compositions, such as the gene compositions described herein. In some embodiments, the vaccine composition contains an antigen that is the target of an antigen-specific adaptive immune response. In other embodiments, the vaccine composition, upon administration, enables the production of an antigen that is the target of an antigen-specific adaptive immune response in an immunized subject. Activation of an adaptive immune response can be detected using methods known in the art, such as measuring the level of antigen-specific antibody production or antigen-specific cell-mediated cytotoxicity.

[0132] Antibody-dependent cell-mediated injury (ADCC) is a form of cytotoxicity in which secretory immunoglobulins bound to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages) enable these cytotoxic effector cells to specifically bind to antigen-carrying target cells, and subsequently kill the target cells with cytotoxins. Antibodies "arm" the cytotoxic cells and are essential for such killing. NK cells, the main cells mediating ADCC, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression in hematopoietic cells is known (see, for example, Ravetch and Kinet, 1991, Annu. Rev. Immunol. 9:457-92). To evaluate the ADCC activity of the target molecule, an in vitro ADCC assay can be performed (see, for example, U.S. Patent Nos. 5,500,362 and 5,821,337). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively or additionally, the ADCC activity of the target molecule may be evaluated in vivo (e.g., in an animal model) (see, for example, Clynes et al., 1998, Proc. Natl. Acad. Sci. USA 95:652-56). Antibodies with little or no ADCC activity may be selected and used.

[0133] Antibody-dependent cell phagocytosis (ADCP) refers to the destruction of target cells via monocyte or macrophage-mediated phagocytosis, where immunoglobulins bound to Fc receptors (FcRs) present on specific phagocytic cells (e.g., neutrophils, monocytes, macrophages) enable these cells to specifically bind to antigen-carrying target cells and subsequently kill them. To evaluate the ADCP activity of a target molecule, an in vitro ADCP assay (e.g., Bracher et al., 2007, J Immunol. Methods 323:160-71) can be performed. Useful phagocytic cells for such assays include peripheral blood mononuclear cells (PBMCs), purified monocytes derived from PBMCs, or U937 cells differentiated into mononuclear cell types. Alternatively or additionally, the ADCP activity of the molecule of interest may be evaluated in vivo (e.g., in an animal model) (see, e.g., Wallace et al., 2001, J. Immunol. Methods 248:167-82). Antibodies with little or no ADCP activity may be selected and used.

[0134] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. An exemplary FcR is a native human FcR. Furthermore, exemplary FcRs are those that bind to IgG antibodies (e.g., gamma receptors) and include the FcγRI, FcγRII, and FcγRIII subclass receptors (including allele variants and alternatively spliced ​​forms of these receptors). FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), which have similar amino acid sequences, primarily differing in their cytoplasmic domains (see, e.g., Daeron, 1997, Annu. Rev. Immunol. 15:203-34). Various FcRs are known (see, for example, Ravetch and Kinet, 1991, Annu.Immunol. 9:457-92; Capel et al., 1994, Immunomethods 4:25-34; and de Haas et al., 1995, J.Lab. Clin.Med. 126:330-41). Other FcRs, including those to be identified in the future, are also included in the term “FcR” as used herein. This term also includes the fetal receptor FcRn, which is involved in the transfer of maternal IgG to the fetus (see, for example, Guyer et al., 1976, J.Immunol. 117:587-93; and Kim et al., 1994, Eu. J.Immunol. 24:2429-34). Antibody variants with improved or reduced binding to FcR have been described (see, for example, WO2000 / 42072; U.S. Patent Nos. 7,183,387; 7,332,581; and 7,335,742; Shields et al. 2001, J. Biol. Biol. Chem. 9(2):6591-604).

[0135] Complement-dependent cell injury, or CDC, refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to an antibody (of the appropriate subclass) bound to a congener antigen. To assess complement activation, a CDC assay (see, e.g., Gazzano-Santoro et al., 1996, J.Immunol. Methods 202:163) can be performed. Polypeptide variants with altered amino acid sequences in the Fc region (polypeptides with variant Fc region) and polypeptide variants with increased or decreased C1q binding ability have been described (see, e.g., U.S. Patent No. 6,194,551; WO1999 / 51642; Idusogie et al., 2000, J.Immunol. 164: 4178-84). Antibodies with little or no CDC activity may be selected and used.

[0136] The term “antibody” is intended to include B cell polypeptide products within the polypeptide immunoglobulin class, which are capable of binding to specific molecular antigens and consist of two identical pairs of polypeptide chains, each pair having one heavy chain (about 50–70 kDa) and one light chain (about 25 kDa), with each amino-terminal portion of each chain containing a variable region of about 100–130 or more amino acids, and each carboxy-terminal portion of each chain containing a constant region. See, for example, Antibody Engineering (Borrebaeck ed., 2d ed. 1995) and Kuby, Immunology (3d ed. 1997). In specific embodiments, specific molecular antigens may be bound by antibodies (including polypeptides, their fragments, or epitopes) provided herein. Antibodies also include, but are not limited to, synthetic antibodies, recombinant antibodies, camelized antibodies, intrabodies, anti-idiotype (anti-Id) antibodies, and any of the functional fragments described above, where a functional fragment refers to a portion of the heavy or light chain polypeptide of an antibody that retains some or all of the binding activity of the antibody from which the fragment originated. Non-limiting examples of functional fragments include single-chain Fv(scFv) (e.g., including monospecific, bispecific, etc.), Fab fragments, F(ab') fragments, F(ab)2 fragments, F(ab')2 fragments, disulfide-bonded Fv(dsFv), Fd fragments, Fv fragments, diabodies, triabodies, tetrabodies, and minibodies. In particular, the antibodies provided herein include immunoglobulin molecules and molecules containing immunologically active portions of immunoglobulin molecules, such as antigen-binding domains or antigen-binding sites (e.g., one or more CDRs of an antibody).Such antibody fragments can be found, for example, in Harlow and Lane, Antibodies: A Laboratory Manual (1989), Mol. Biology and Biotechnology: A Comprehensive Desk Reference (Myers ed., 1995), Huston et al., 1993, Cell Biophysics 22:189-224, Pluckthun and Skerra, 1989, Meth. Enzymol. Enzymol. 178:497-515, and Day, Advanced Immunochemistry (2nd ed. 1990). The antibodies provided herein may be of any class of immunoglobulin molecule (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2).

[0137] The terms “administer” or “dosage” refer to the act of injecting or otherwise physically delivering an extracorporeal substance (e.g., the lipid nanoparticle composition described herein) to a patient by mucosal, intradermal, intravenous, intramuscular delivery, and / or any other physical delivery method described herein or known in the art. Where a disease, disorder, condition, or its symptoms is being treated, the administration of the substance is typically performed after the onset of the disease, disorder, condition, or its symptoms. Where a disease, disorder, condition, or its symptoms is being prevented, the administration of the substance is typically performed before the onset of the disease, disorder, condition, or its symptoms.

[0138] "Chronic" administration refers to the continuous administration of a drug(s) (for example, over a period of several days, weeks, months, or years) to maintain the initial therapeutic effect (activity) for an extended period, as opposed to an acute mode. "Intermittent" administration refers to treatment that is not performed continuously without interruption, but is effectively cyclical.

[0139] As used herein, the terms “targeted delivery” or the verb form “target” refer to a process that facilitates the delivery of a drug (such as a therapeutic payload molecule in a lipid nanoparticle composition described herein) to reach a specific organ, tissue, cell, and / or intracellular compartment (referred to as a targeted site) more than any other organ, tissue, cell, or intracellular compartment (referred to as a non-targeted site). Targeted delivery can be detected using methods known in the art, for example, by comparing the concentration of the delivered drug in a targeted cell population after systemic administration to the concentration of the delivered drug in a non-targeted cell population. In certain embodiments, targeted delivery results in a concentration at the targeted site that is at least twice as high as at the non-targeted site.

[0140] An "effective dose" is generally defined as an amount sufficient to reduce the severity and / or frequency of symptoms, eliminate symptoms and / or their underlying causes, prevent the onset of symptoms and / or their underlying causes, and / or improve or correct damage resulting from or associated with a disease, disorder, or condition, including, for example, infection and tumors. In some embodiments, the effective dose is a therapeutic effective dose or a preventive effective dose.

[0141] As used herein, the term “therapeutic dose” means an amount of an agent (e.g., a vaccine composition) sufficient to reduce and / or improve the severity and / or duration of a given disease, disorder, or condition and / or symptoms associated therewith (e.g., an infectious disease such as one caused by a viral infection, or a neoplastic disease such as cancer). The “therapeutic dose” of a substance / molecule / agent of this disclosure (e.g., a lipid nanoparticle composition described herein) may vary depending on factors such as the individual’s medical condition, age, sex, and weight, as well as the ability of the substance / molecule / agent to induce a desired response in the individual. The therapeutic dose encompasses an amount in which the therapeutically beneficial effect outweighs any toxic or adverse effects of the substance / molecule / agent. In certain embodiments, the term “therapeutic dose” means an amount of a lipid nanoparticle composition described herein, or a therapeutic or prophylactic agent (e.g., therapeutic mRNA) contained therein, that is effective in “treating” a disease, disorder, or condition in a subject or mammal.

[0142] "Preventive dose" refers to the amount of a drug or pharmaceutical composition (e.g., a vaccine composition) administered to a subject that, when administered, has the intended preventive effect (e.g., prevention, delay, or reduction of the likelihood of the onset (or recurrence) of a disease, disorder, condition, or associated symptom(s) (e.g., an infectious disease caused by a viral infection, or a neoplastic disease such as cancer)). In some embodiments, the term "preventive dose" refers to the amount of the lipid nanoparticle composition described herein, or the preventive agent contained therein (e.g., nucleic acid, e.g., nucleic acid (including mRNA) as described in Section 5.4 or 6), that is effective in "preventing" a disease, disorder, or condition in a subject or mammal. Typically, but not necessarily, the preventive dose may be less than the therapeutic dose because the preventive dose is used on a subject before or at an early stage of the disease, disorder, or condition. Complete therapeutic or preventive effects do not necessarily occur with a single dose and may only occur after a series of doses have been administered. Therefore, a therapeutic or preventive dose may be administered in one or more doses.

[0143] The terms “prevent,” “prevention,” and “prevention” refer to reducing the likelihood of the onset (or recurrence) of a disease, disorder, condition, or associated symptoms (for example, an infectious disease (caused by a viral infection), or a neoplastic disease such as cancer).

[0144] The terms “to manage,” “to control,” and “manage” refer to the beneficial effects that a subject receives from a treatment (e.g., a preventive or therapeutic agent) and do not result in a cure for the disease. In certain embodiments, a subject is administered one or more therapies (e.g., a preventive or therapeutic agent such as the lipid nanoparticle composition described herein) to “manage” an infectious or neoplastic disease, one or more of their symptoms, and to prevent the progression or worsening of the disease.

[0145] The term “preventive agent” refers to any agent that can, in whole or in part, inhibit the onset, recurrence, onset, or progression of a disease and / or related symptoms in a subject. In some embodiments, the preventive agent comprises or consists of nucleic acids described herein (e.g., nucleic acids described in Section 5.4 or Section 6). In some embodiments, the preventive agent comprises or consists of proteins described herein (e.g., proteins described in Section 5.3). In some embodiments, the preventive agent comprises or consists of a vector containing nucleic acids described herein.

[0146] The term “therapeutic agent” refers to any agent that can be used to treat, prevent, or alleviate a disease, disorder, or condition (including the treatment, prevention, or alleviation of one or more symptoms and / or symptoms associated therewith of a disease, disorder, or condition). In some embodiments, the therapeutic agent is a nucleic acid as described herein (e.g., a nucleic acid as described in Section 5.4 or Section 6). In some embodiments, the therapeutic agent is a protein as described herein (e.g., a protein as described in Section 5.3). In some embodiments, the preventive agent comprises or consists of a vector containing a nucleic acid as described herein.

[0147] The term “therapy” refers to any protocol, method, and / or agent that can be used to prevent, manage, treat, and / or improve a disease, disorder, or condition. In certain embodiments, the terms “therapy(plural)” and “therapy(singular)” refer to biological therapies, supportive therapies, and / or other therapies known to those skilled in the art, such as healthcare professionals, that are useful for preventing, managing, treating, and / or improving a disease, disorder, or condition.

[0148] As used herein, “prophylactically effective serum titer” is the serum titer of an antibody in a subject (e.g., a human) that completely or partially inhibits the onset, recurrence, development, or progression of a disease, disorder, or condition and / or symptoms associated therewith in the subject.

[0149] In a particular embodiment, “therapeutic serum titer” is the serum titer of an antibody in a subject (e.g., a human) that reduces the severity, duration, and / or symptoms associated with a disease, disorder, or condition in the subject.

[0150] The term "serum titer" refers to the average serum titer from multiple samples (e.g., at multiple time points) in a subject, or in a population of at least 10, at least 20, at least 40 subjects, up to approximately 100, 1000, or more.

[0151] The term “adverse effects” encompasses undesirable and / or harmful effects of a therapy (e.g., a preventive or therapeutic agent). Undesirable effects are not necessarily harmful. Adverse effects from a therapy (e.g., a preventive or therapeutic agent) may be harmful, unpleasant, or dangerous. Examples of adverse effects include diarrhea, cough, gastroenteritis, wheezing, nausea, vomiting, anorexia, abdominal cramps, fever, pain, weight loss, dehydration, hair loss, difficulty breathing, insomnia, dizziness, mucositis, nerve and muscle effects, fatigue, dry mouth, loss of appetite, rash or swelling at the injection site, flu-like symptoms such as fever, chills, and fatigue, digestive problems, and allergic reactions. There are many other undesirable effects that patients may experience, and these are known in the art. Many are described in Physician's Desk Reference (68th ed. 2014).

[0152] The terms “subject” and “patient” may be used interchangeably. As used herein, in certain embodiments, the subject is a mammal such as a non-primate (e.g., cattle, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkeys and humans). In specific embodiments, the subject is a human. In one embodiment, the subject is a mammal (e.g., a human) with an infectious or neoplastic disease. In another embodiment, the subject is a mammal (e.g., a human) at risk of developing an infectious or neoplastic disease.

[0153] The term "elderly person" refers to a person aged 65 or older. The term "adult person" refers to a person aged 18 or older. The term "child person" refers to a person aged 1 to 18 years. The term "infant person" refers to a person aged 1 to 3 years. The term "newborn person" refers to a person aged 1 year or younger.

[0154] A “detectable probe” refers to a composition that provides a detectable signal. This term includes, but is not limited to, any fluorophore, chromophore, radiolabel, enzyme, antibody, or antibody fragment that provides a detectable signal through its activity.

[0155] The term "detectable agent" refers to a substance that can be used to confirm the existence or presence of a desired molecule, such as an antigen encoded by an mRNA molecule described herein, in a sample or subject. A detectable agent may be a substance that can be visualized or otherwise determined and / or measured (e.g., by quantification).

[0156] "Substantially all" means at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or about 100%.

[0157] As used herein, and unless otherwise indicated, the terms “about” or “approximately” mean a tolerance of a particular value as determined by those skilled in the art, which depends on how the value is measured or determined. In certain embodiments, the terms “about” or “approximately” mean within one, two, three, or four standard deviations. In certain embodiments, the terms “about” or “approximately” mean within 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.05%, or less of a given value or range.

[0158] Unless otherwise explicitly indicated by the context, the singular terms "a," "an," and "the" as used herein refer to multiple objects.

[0159] All publications, patent applications, accession numbers, and other references cited herein are incorporated herein by reference in whole, as if each individual publication or patent application were explicitly and individually indicated as being invoked by reference. Publications discussed herein refer only to disclosures prior to the filing date of this application. Nothing in this specification constitutes an admission that the present invention has no prior rights to such publications based on prior inventions. Furthermore, the publication dates provided may differ from the actual publication dates, which may need to be verified on a case-by-case basis.

[0160] Numerous embodiments of the present invention are described. However, it will be understood that various modifications may be made without departing from the spirit and scope of this disclosure. Accordingly, the descriptions in the experimental section and the examples are intended to illustrate (but not limit) the scope of the invention as described in the claims.

[0161] 5.3 Protein VZV glycoproteins include glycoprotein E (gE), glycoprotein B (gB), glycoprotein H (gH), and glycoprotein L (gL), with gE being the most abundantly expressed in VZV-infected cells. VZV gE is generally 623 amino acid residues long and contains a signal peptide (generally amino acid residues 1-30 of full-length gE; for example, the signal peptide of GenBank accession number AAG32558.1 consists of amino acid residues 1-30 of GenBank accession number AAG32558.1) and a transmembrane domain. Mature gE lacks the signal peptide. VZV gE forms a heterodimer with gI, and this heterodimer gE / gI is required for intercellular diffusion of the virus. Furthermore, the VZV gE / gI heterodimer interacts with the Fc region of IgG. VZV gE binds to insulin-degrading enzyme (IDE). VZV gE is phosphorylated by the viral kinase encoded by ORF47. Exemplary VZV gE can be found in GenBank accession numbers AAG32558.1, AAF61669.1, AAK19946.1, AAK01056.1, AAK19955.1, ABF21641.1, ABF22006.1, ABF22152.1, ABF22152.1, ABF22225.1, ABF22298.1, AEW88044.1, AEW88116.1, AAY57677.1, AAY57748.1, Q9J3M8.1, CAA27951.1, QCA47220.1, AEW88980.1, WWU03079.1, and AEW88548.1, as well as Uniprot numbers Q9J3M8 and P09259. In some embodiments, VZV gE is gE from the Dumas strain. In some embodiments, VZV gE is gE from the KPZ13-287 strain. In some embodiments, VZV gE is gE from the VZVi / Munich.GER / 30.07 / Z[3] strain. In some embodiments, VZV gE is gE from the NSYY3 strain. In some embodiments, VZV gE is gE from the 1002 / 2008 strain. In some embodiments, VZV gE is gE from the NSYY3 strain. In some embodiments, VZV gE is gE from the Oka strain.

[0162] In some embodiments, what is provided herein is a fragment of mature VZV gE, the fragment comprising a cleavage of at least one amino acid residue, up to 50, 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage comprises a cleavage of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of 11, 12, 13, 14, 15, 16, 17, 18, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of up to 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of 14 or 37 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of 37 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of up to 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, or 31 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is the cleavage of up to 30 or 29 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is a cleavage of up to 28 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is a cleavage of up to 27 or 26 amino acid residues from the C-terminus of mature gE. In some embodiments, the cleavage is a cleavage of up to 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2 amino acid residues from the C-terminus of mature gE, or up to 1 amino acid residue. In some embodiments, the mature gE contains the amino acid sequence described in SEQ ID NO: 1.

[0163] In some embodiments, the mature gE fragment further comprises one or more amino acid substitutions. In some embodiments, the mature gE fragment further comprises one, two, three, four, five, or all of the amino acid substitutions selected from Y569A, Y582G, S593A, S595A, T596A, and T598A, where amino acid residue positions 569, 582, 593, 595, 596, and 598 are amino acid residue position numbers according to the full-length VZV gE. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in Sequence ID No. 55.

[0164] In some embodiments, the mature gE fragment further comprises the amino acid residue substitution Y569A, where amino acid residue position number 569 is the amino acid residue position number according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises the amino acid residue substitution Y582G, where amino acid residue position 582 is the amino acid residue position number according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises the amino acid residue substitutions Y569A and Y582G, where amino acid residue position numbers 569 and 582 are the amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises the amino acid residue substitutions Y569A, Y582G, and S593A, where amino acid residue position numbers 569, 582, and 593 are the amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, Y582G, and S595A, where amino acid residue positions 569, 582, and 595 are amino acid residue positions according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, Y582G, and T596A, where amino acid residue positions 569, 582, and 596 are amino acid residue positions according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, Y582G, and T598A, where amino acid residue positions 569, 582, and 598 are amino acid residue positions according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, Y582G, S593A, and S595A, where amino acid residue position numbers 569, 582, 593, and 595 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, Y582G, S593A, S595A, and T596A, where amino acid residue position numbers 569, 582, 593, 595, and 596 are amino acid residue position numbers according to full-length VZV gE.In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A, where amino acid residue position numbers 569, 582, 593, 595, 596, and 598 are amino acid residue position numbers according to the full-length VZV gE. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in Sequence ID No. 55.

[0165] In some embodiments, the mature gE fragment further comprises amino acid residue substitution S593A, where amino acid residue position 593 is the amino acid residue position number according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitution S595A, where amino acid residue position 595 is the amino acid residue position number according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitution T596A, where amino acid residue position 596 is the amino acid residue position number according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitution T598A, where amino acid residue position 598 is the amino acid residue position number according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S593A and S595A, where amino acid residue positions 593 and 595 are the amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S593A and T596A, where amino acid residue positions 593 and 596 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S593A and T598A, where amino acid residue positions 593 and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S595A and T596A, where amino acid residue positions 595 and 596 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S595A and T598A, where amino acid residue positions 595 and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions T596A and T598A, where amino acid residue positions 596 and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S593A, S595A, and T596A, where amino acid residue positions 593, 595, and 596 are amino acid residue position numbers according to full-length VZV gE.In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S593A, S595A, and T598A, where amino acid residue positions 593, 595, and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S595A, T596A, and T598A, where amino acid residue positions 595, 596, and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions S593A, S595A, T596A, and T598A, where amino acid residue positions 593, 595, 596, and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A, where amino acid residue position numbers 582, 593, 595, 596, and 598 are amino acid residue position numbers according to the full-length VZV gE. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in Sequence ID No. 55.

[0166] In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A and S593A, where amino acid residue positions 569 and 593 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A and S595A, where amino acid residue positions 569 and 595 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A and T596A, where amino acid residue positions 569 and 596 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A and T598A, where amino acid residue positions 569 and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S593A, and S595A, where amino acid residue positions 569, 593, and 595 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S593A, and T596A, where amino acid residue positions 569, 593, and 596 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S593A, and T598A, where amino acid residue positions 569, 593, and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S595A, and T596A, where amino acid residue positions 569, 595, and 596 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S595A, and T598A, where amino acid residue positions 569, 595, and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, T596A, and T598A, where amino acid residue positions 569, 596, and 598 are amino acid residue position numbers according to full-length VZV gE.In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S593A, S595A, and T596A, where amino acid residue positions 569, 593, 595, and 596 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S593A, S595A, and T598A, where amino acid residue positions 569, 593, 595, and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S595A, T596A, and T598A, where amino acid residue positions 569, 595, 596, and 598 are amino acid residue position numbers according to full-length VZV gE. In some embodiments, the mature gE fragment further comprises amino acid residue substitutions Y569A, S593A, S595A, T596A, T598A, where amino acid residue positions 569, 593, 595, 596, and 598 are amino acid residue position numbers according to the full-length VZV gE. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in Sequence ID No. 55.

[0167] In some embodiments, provided herein is a fusion protein comprising a fragment of mature gE of VZV and a heterologous signal peptide, wherein the N-terminus of the fragment is fused to the C-terminus of the heterologous signal peptide. In some embodiments, the mature gE comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the heterologous signal peptide is a human tPA signal peptide. In some embodiments, the human tPA signal peptide comprises the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the human tPA signal peptide comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the human tPA signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the heterologous signal peptide is a human IgE signal peptide. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the mature gE fragment is a fragment described herein (for example, a mature gE fragment described in the examples in this section or Section 6). In some embodiments, the mature gE fragment comprises the amino acid sequence of SEQ ID NO: 3, 6, 8, 10, or 12. In a specific embodiment, the fragment comprises the amino acid sequence of SEQ ID NO: 6. In some embodiments, the mature gE fragment comprises an amino acid sequence that is at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identical to SEQ ID NO: 3, 6, 8, 10, or 12.In some embodiments, the mature gE fragment contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to SEQ ID NOs. 3, 6, 8, 10, or 12. In some embodiments, the mature gE fragment contains an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to SEQ ID NOs. 3, 6, 8, 10, or 12. In some embodiments, the mature gE fragment contains an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NOs. 3, 6, 8, 10, or 12.

[0168] In some embodiments, provided herein is a fusion protein comprising a fragment of mature VZV gE and a human IgE signal peptide, wherein the fragment comprises a cleavage of 37 amino acid residues from the C-terminus of mature gE, and the N-terminus of the fragment is fused to the C-terminus of the human IgE signal peptide. In some embodiments, provided herein is a fusion protein comprising a fragment of mature VZV gE and a human IgE signal peptide, wherein the fragment comprises a cleavage of 37 amino acid residues from the C-terminus of mature gE, as well as amino acid residue substitutions Y569A and Y582G, where amino acid residue positions 569 and 582 are amino acid residue position numbers according to full-length VZV gE, and the N-terminus of the fragment is fused to the C-terminus of the human IgE signal peptide. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23.

[0169] In some embodiments, provided herein is a fusion protein comprising a fragment of mature VZV gE and a human IgE signal peptide, the fragment comprising a cleavage of 37 amino acid residues from the C-terminus of mature gE, as well as amino acid residue substitutions Y569A and Y582G, where amino acid residues 569 and 582 are positional numbers of amino acid residues according to full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the fragment comprises the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the fusion protein includes an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence of SEQ ID NO: 59. In some embodiments, the fusion protein includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 59.

[0170] In some embodiments, provided herein is a fusion protein comprising a variant of mature VZV gE and a human IgE signal peptide, the variant comprising amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A, where amino acid residues 569, 582, 593, 595, 596, and 598 are positional numbers of amino acid residues according to full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence of SEQ ID NO: 55. In some embodiments, the variant comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23.

[0171] In some embodiments, provided herein is a fusion protein comprising a variant of mature VZV gE and a human IgE signal peptide, wherein the variant comprises amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A, where amino acid residues 582, 593, 595, 596, and 598 are positional numbers of amino acid residues according to full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the variant comprises the amino acid sequence described in SEQ ID NO: 12. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23.

[0172] In some embodiments, provided herein is a fusion protein comprising a fragment of mature VZV gE and a human IgE signal peptide, the fragment comprising a 50-amino acid cleavage from the C-terminus of mature gE, as well as an amino acid residue substitution Y569A, where amino acid residue 569 is the position number of an amino acid residue according to full-length VZV gE. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the fragment comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the human IgE signal peptide comprises the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23.

[0173] In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising amino acid residue substitutions Y569A and Y582G relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising amino acid residue substitutions Y569A, Y582G, and S593A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising amino acid residue substitutions Y569A, Y582G, and S595A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising amino acid residue substitutions Y569A, Y582G, and T596A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising amino acid residue substitutions Y569A, Y582G, and T598A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising amino acid residue substitutions Y569A, Y582G, S593A, and S595A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising amino acid residue substitutions Y569A, Y582G, S593A, and T596A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising amino acid residue substitutions Y569A, Y582G, S593A, and T598A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising the amino acid residue substitutions Y569A, Y582G, S595A, and T596A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising the amino acid residue substitutions Y569A, Y582G, S595A, and T598A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising the amino acid residue substitutions Y569A, Y582G, T596A, and T598A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising the amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A relative to full-length VSV gE. In some embodiments, provided herein are full-length gE variants of VZV, the variants comprising the amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A compared to full-length VSV gE. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in Sequence ID No. 55.

[0174] In some embodiments, the herein provides a protein comprising a variant of mature VZV gE, the variant comprising (i) a cleavage of 37 amino acid residues from the C-terminus of mature gE, and (ii) amino acid residue substitutions Y569A and Y582G, where amino acid residue positions 569 and 582 are the amino acid residue position numbers of full-length VSV gE. In some embodiments, mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the variant comprises the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the amino acid sequence of the variant comprises the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the variant comprises an amino acid sequence that is at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identical to the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the variant comprises an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the variant includes an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the variant includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the protein further includes a signal peptide of VZV gE. In some embodiments, the signal peptide of VZV gE includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the protein further includes a heterologous signal peptide, the N-terminus of the variant is fused to the C-terminus of the heterologous signal peptide.In some embodiments, the heterologous signal peptide is a human IgE signal peptide. In some embodiments, the human IgE signal peptide includes the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 6, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the protein includes the amino acid sequence described in SEQ ID NO: 59. In some embodiments, the amino acid sequence of the protein consists of the amino acid sequence described in SEQ ID NO: 59. In some embodiments, the heterologous signal peptide of the protein is a human tPA signal peptide. In some embodiments, the human tPA signal peptide includes the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the human tPA signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 28.

[0175] In some embodiments, the herein provides a protein comprising a variant of mature VZV gE, the variant comprising amino acid residue substitutions Y569A and Y582G, where amino acid residue positions 569 and 582 are the amino acid residue position numbers of full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the variant comprises the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the variant comprises an amino acid sequence that is at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identical to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the variant comprises an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the variant includes an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the variant includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the protein further includes a VZV gE signal peptide. In some embodiments, the VZV gE signal peptide includes the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the VZV gE signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the protein further includes a heterologous signal peptide, the N-terminus of the variant fused to the C-terminus of the heterologous signal peptide. In some embodiments, the heterologous signal peptide is a human IgE signal peptide.In some embodiments, the human IgE signal peptide includes the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 8, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the heterologous signal peptide of the protein is a human tPA signal peptide. In some embodiments, the human tPA signal peptide includes the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the human tPA signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 28.

[0176] In some embodiments, the herein provides a protein comprising a variant of mature VZV gE, the variant comprising amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A, where amino acid residue positions 569, 582, 593, 595, 596, and 598 are the amino acid residue position numbers of full-length VSV gE. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the variant comprises the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the variant comprises an amino acid sequence that is at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identical to the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the variant includes an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the variant includes an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the variant includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the protein further includes a signal peptide of VZV gE. In some embodiments, the signal peptide of VZV gE includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the protein further contains a heterologous signal peptide, where the N-terminus of the variant is fused to the C-terminus of the heterologous signal peptide.In some embodiments, the heterologous signal peptide is a human IgE signal peptide. In some embodiments, the human IgE signal peptide includes the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 10, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the heterologous signal peptide of the protein is a human tPA signal peptide. In some embodiments, the human tPA signal peptide includes the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the human tPA signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 28.

[0177] In some embodiments, the herein provides a protein comprising a variant of mature VZV gE, the variant comprising amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A, where amino acid residue positions 582, 593, 595, 596, and 598 are the amino acid residue position numbers of full-length VSV gE. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the variant comprises the amino acid sequence described in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 12. In some embodiments, the variant comprises an amino acid sequence that is at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identical to the amino acid sequence described in SEQ ID NO: 12. In some embodiments, the variant includes an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to the amino acid sequence described in SEQ ID NO: 12. In some embodiments, the variant includes an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 12. In some embodiments, the variant includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 12. In some embodiments, the protein further includes a signal peptide of VZV gE. In some embodiments, the signal peptide of VZV gE includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the protein further contains a heterologous signal peptide, where the N-terminus of the variant is fused to the C-terminus of the heterologous signal peptide.In some embodiments, the heterologous signal peptide is a human IgE signal peptide. In some embodiments, the human IgE signal peptide includes the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 12, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the heterologous signal peptide of the protein is a human tPA signal peptide. In some embodiments, the human tPA signal peptide includes the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the human tPA signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 28.

[0178] In some embodiments, the herein provides a protein comprising a variant of mature VZV gE, the variant comprising (i) a cleavage of 50 amino acid residues from the C-terminus of the mature gE protein, and (ii) an amino acid residue substitution Y569A, where amino acid residue position 569 is the amino acid residue position of full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO: 55. In some embodiments, the variant comprises the amino acid sequence described in SEQ ID NO: 3. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 3. In some embodiments, the variant comprises an amino acid sequence that is at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identical to the amino acid sequence described in SEQ ID NO: 3. In some embodiments, the variant comprises an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to the amino acid sequence described in SEQ ID NO: 3. In some embodiments, the variant contains an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the amino acid sequence described in SEQ ID NO: 3. In some embodiments, the variant contains an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 3. In some embodiments, the protein further contains a VZV gE signal peptide containing the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the VZV gE signal peptide contains an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 18. In some embodiments, the protein further contains a heterologous signal peptide, the N-terminus of the variant fused to the C-terminus of the heterologous signal peptide. In some embodiments, the heterologous signal peptide is a human IgE signal peptide.In some embodiments, the human IgE signal peptide includes the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the variant consists of the amino acid sequence described in SEQ ID NO: 3, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO: 23. In some embodiments, the heterologous signal peptide of the protein is a human tPA signal peptide. In some embodiments, the human tPA signal peptide includes the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the human tPA signal peptide includes an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO: 28.

[0179] In some embodiments, the protein provided herein contains the amino acid sequence of VZV gE mutain disclosed in Table 1. In some embodiments, the protein provided herein contains the amino acid sequence of VZV gE mutain-1 disclosed in Table 1. In some embodiments, the protein provided herein contains the amino acid sequence of VZV gE mutain-2 disclosed in Table 1. In some embodiments, the protein provided herein contains the amino acid sequence of VZV gE mutain-3 disclosed in Table 1. In some embodiments, the protein provided herein contains the amino acid sequence of VZV gE mutain-4 disclosed in Table 1. In some embodiments, the protein provided herein contains the amino acid sequence of VZV gE mutain-5 disclosed in Table 1.

[0180] In some embodiments, what is provided herein are variants of the VZV gE protein described in Section 6 below. In some embodiments, what is provided herein are variants of VZV gE other than the control described in Section 6 below. In some embodiments, what is provided herein are variants of VZV gE encoded by nucleic acids other than the control described in Section 6 below.

[0181] In some embodiments, the fragments, variants, or fusion proteins described herein have a structure similar to wild-type VZV gE, as assessed by techniques known to those skilled in the art, such as X-ray crystallography, nuclear magnetic resonance, or binding to an antibody specific to the structural epitope of wild-type VZV gE. In some embodiments, the proteins described herein, including variants of mature gE, have a structure similar to wild-type VZV gE, as assessed by techniques known to those skilled in the art, such as X-ray crystallography, nuclear magnetic resonance, or binding to an antibody specific to the structural epitope of wild-type VZV gE.

[0182] In some embodiments, the fragments, variants, or fusion proteins described herein retain at least one activity or function of mature VZV gE. In some embodiments, the proteins described herein, comprising variants of mature gE, retain at least one activity or function of mature VZV gE. For example, in some embodiments, the ability to form heterodimers with gI is retained. In other embodiments, the ability to bind to the Fc receptor is retained. In other embodiments, the ability to bind to insulin-degrading enzymes (IDEs) is retained. In other embodiments, the ability to be phosphorylated by the viral kinase encoded by ORF47 is retained.

[0183] 5.4 Therapeutic Nucleic Acids In one embodiment, the herein provides a therapeutic nucleic acid molecule for the management, prevention, and treatment of VZV infection. In some embodiments, the therapeutic nucleic acid encodes a peptide or polypeptide, which, when administered to a target requiring it, is expressed by cells within the target and produces the encoded peptide or polypeptide. In some embodiments, the therapeutic nucleic acid molecule is a DNA molecule. In other embodiments, the therapeutic nucleic acid molecule is an RNA molecule. In certain embodiments, the therapeutic nucleic acid molecule is an mRNA molecule.

[0184] In some embodiments, the therapeutic nucleic acid molecule is formulated in a vaccine composition. In some embodiments, the vaccine composition is a gene vaccine as described herein. In some embodiments, the vaccine composition comprises an mRNA molecule as described herein.

[0185] In some embodiments, the mRNA molecules of this disclosure encode a peptide or polypeptide of interest (including any naturally occurring or non-naturally occurring or otherwise modified polypeptide). The peptide or polypeptide encoded by the mRNA may be of any size and may have any secondary structure or activity. In some embodiments, the polypeptide encoded by the mRNA payload may have therapeutic effects when expressed in cells.

[0186] In some embodiments, the mRNA molecule of this disclosure includes at least one coding region (e.g., an open reading frame (ORF)) encoding the peptide or polypeptide of interest. In some embodiments, the nucleic acid molecule further includes at least one untranslated region (UTR). In certain embodiments, the untranslated region (UTR) is located upstream (towards the 5' end) of the coding region and is referred to herein as the 5'-UTR. In certain embodiments, the untranslated region (UTR) is located downstream (towards the 3' end) of the coding region and is referred to herein as the 3'-UTR. In certain embodiments, the nucleic acid molecule includes both the 5'-UTR and the 3'-UTR. In some embodiments, the 5'-UTR includes a 5' cap structure. In some embodiments, the nucleic acid molecule includes a Kozak sequence (e.g., in the 5'-UTR). In some embodiments, the nucleic acid molecule includes a poly-A region (e.g., in the 3'-UTR). In some embodiments, the nucleic acid molecule includes a polyadenylation signal (e.g., in the 3'-UTR). In some embodiments, the nucleic acid molecule includes a stabilizing region (e.g., in the 3'-UTR). In some embodiments, the nucleic acid molecule includes a secondary structure. In some embodiments, the secondary structure is a stem-loop. In some embodiments, the nucleic acid molecule includes a stem-loop sequence (e.g., in the 5'-UTR and / or 3'-UTR). In some embodiments, the nucleic acid molecule includes one or more intron regions that can be excised during splicing. In specific embodiments, the nucleic acid molecule includes one or more regions selected from the 5'-UTR and the coding region. In specific embodiments, the nucleic acid molecule includes one or more regions selected from the coding region and the 3'-UTR. In specific embodiments, the nucleic acid molecule includes one or more regions selected from the 5'-UTR, the coding region, and the 3'-UTR.

[0187] 5.4.1 Code Domain In some embodiments, the nucleic acid molecules of the present disclosure include at least one coding region. In some embodiments, the coding region is an open reading frame (ORF) encoding a single peptide or protein. In some embodiments, the coding region includes at least two ORFs, each encoding a peptide or protein. In those embodiments, where the coding region includes two or more ORFs, the encoded peptides and / or proteins may be the same as or different from each other. In some embodiments, multiple ORFs within the coding region are separated by non-coding sequences. In specific embodiments, the non-coding sequences separating two ORFs include an internal ribosome entry site (IRES).

[0188] While not theoretically bound, it is intended that internal ribosome entry sites (IRESs) can function as either a single ribosome binding site or as one of several ribosome binding sites on mRNA. mRNA molecules containing multiple functional ribosome binding sites can encode several peptides or proteins that are independently translated by ribosomes (e.g., multicistronic mRNA). Therefore, in some embodiments, the nucleic acid molecules (e.g., mRNA) of this disclosure contain one or more internal ribosome entry sites (IRESs). Examples of IRES sequences that can be used in connection with this disclosure include, but are not limited to, those derived from picomaviruses (e.g., FMDV), plague virus (CFFV), poliovirus (PV), encephalomyocarditis virus (ECMV), foot-and-mouth disease virus (FMDV), hepatitis C virus (HCV), swine cholera virus (CSFV), mouse leukemia virus (MLV), simian immunodeficiency virus (SIV), or cricket paralysis virus (CrPV).

[0189] In various embodiments, the nucleic acid molecules of the Disclosure encode at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10 peptides or proteins. The peptides and proteins encoded by the nucleic acid molecules may be the same or different. In some embodiments, the nucleic acid molecules of the Disclosure encode dipeptides (e.g., camosine and anserine). In some embodiments, the nucleic acid molecules encode tripeptides. In some embodiments, the nucleic acid molecules encode tetrapeptides. In some embodiments, the nucleic acid molecules encode pentapeptides. In some embodiments, the nucleic acid molecules encode hexapeptides. In some embodiments, the nucleic acid molecules encode heptapeptides. In some embodiments, the nucleic acid molecules encode octapeptides. In some embodiments, the nucleic acid molecules encode nonapeptides. In some embodiments, the nucleic acid molecules encode decapeptides. In some embodiments, the nucleic acid molecules encode peptides or polypeptides having at least about 15 amino acids. In some embodiments, the nucleic acid molecules encode peptides or polypeptides having at least about 50 amino acids. In some embodiments, the nucleic acid molecules encode peptides or polypeptides having at least about 100 amino acids. In some embodiments, the nucleic acid molecule encodes a peptide or polypeptide having at least about 150 amino acids. In some embodiments, the nucleic acid molecule encodes a peptide or polypeptide having at least about 300 amino acids. In some embodiments, the nucleic acid molecule encodes a peptide or polypeptide having at least about 500 amino acids. In some embodiments, the nucleic acid molecule encodes a peptide or polypeptide having at least about 1000 amino acids.

[0190] In some embodiments, the nucleic acid molecule of the Disclosure is at least about 30 nucleotides (nt) long. In some embodiments, the nucleic acid molecule is at least about 35 nt long. In some embodiments, the nucleic acid molecule is at least about 40 nt long. In some embodiments, the nucleic acid molecule is at least about 45 nt long. In some embodiments, the nucleic acid molecule is at least about 50 nt long. In some embodiments, the nucleic acid molecule is at least about 55 nt long. In some embodiments, the nucleic acid molecule is at least about 60 nt long. In some embodiments, the nucleic acid molecule is at least about 65 nt long. In some embodiments, the nucleic acid molecule is at least about 70 nt long. In some embodiments, the nucleic acid molecule is at least about 75 nt long. In some embodiments, the nucleic acid molecule is at least about 80 nt long. In some embodiments, the nucleic acid molecule is at least about 85 nt long. In some embodiments, the nucleic acid molecule is at least about 90 nt long. In some embodiments, the nucleic acid molecule is at least about 95 nt long. In some embodiments, the nucleic acid molecule is at least about 100 nt long. In some embodiments, the nucleic acid molecule is at least about 120 nt in length. In some embodiments, the nucleic acid molecule is at least about 140 nt in length. In some embodiments, the nucleic acid molecule is at least about 160 nt in length. In some embodiments, the nucleic acid molecule is at least about 180 nt in length. In some embodiments, the nucleic acid molecule is at least about 200 nt in length. In some embodiments, the nucleic acid molecule is at least about 250 nt in length. In some embodiments, the nucleic acid molecule is at least about 300 nt in length. In some embodiments, the nucleic acid molecule is at least about 400 nt in length. In some embodiments, the nucleic acid molecule is at least about 500 nt in length. In some embodiments, the nucleic acid molecule is at least about 600 nt in length. In some embodiments, the nucleic acid molecule is at least about 700 nt in length. In some embodiments, the nucleic acid molecule is at least about 800 nt in length. In some embodiments, the nucleic acid molecule is at least about 900 nt in length. In some embodiments, the nucleic acid molecule is at least about 1000 nt in length.In some embodiments, the nucleic acid molecule is at least about 1100 nt long. In some embodiments, the nucleic acid molecule is at least about 1200 nt long. In some embodiments, the nucleic acid molecule is at least about 1300 nt long. In some embodiments, the nucleic acid molecule is at least about 1400 nt long. In some embodiments, the nucleic acid molecule is at least about 1500 nt long. In some embodiments, the nucleic acid molecule is at least about 1600 nt long. In some embodiments, the nucleic acid molecule is at least about 1700 nt long. In some embodiments, the nucleic acid molecule is at least about 1800 nt long. In some embodiments, the nucleic acid molecule is at least about 1900 nt long. In some embodiments, the nucleic acid molecule is at least about 2000 nt long. In some embodiments, the nucleic acid molecule is at least about 2500 nt long. In some embodiments, the nucleic acid molecule is at least about 3000 nt long. In some embodiments, the nucleic acid molecule is at least about 3500 nt long. In some embodiments, the nucleic acid molecule is at least about 4000 nt long. In some embodiments, the nucleic acid molecule is at least about 4500 nt in length. In some embodiments, the nucleic acid molecule is at least about 5000 nt in length.

[0191] In specific embodiments, the therapeutic nucleic acids of the present disclosure are formulated as vaccine compositions described herein (e.g., gene vaccines). In some embodiments, the therapeutic nucleic acids encode peptides or proteins capable of inducing immunity against one or more target conditions or diseases. In some embodiments, the target conditions are associated with or caused by infection with a pathogen such as VZV. In some embodiments, the therapeutic nucleic acid sequence (e.g., mRNA) encodes a pathogen-specific pathogenic protein or immunogenic fragment (e.g., an epitope) or a derivative thereof. When administered to a target for vaccination, the vaccine enables the expression of the encoded pathogenic protein (or immunogenic fragment or derivative thereof), thereby inducing immunity against the pathogen in the target.

[0192] In specific embodiments, provided herein are therapeutic compositions (e.g., vaccine compositions) for the management, prevention, and treatment of diseases or disorders caused by or resulting from VZV.

[0193] While not bound by theory, VZV, or varicella-zoster virus (also known as human herpesvirus 3), is assumed to be a double-stranded DNA virus belonging to the alphaherpesvirus group. VZV has only one serotype. The VZV genome contains 71 genes and codes for 67 proteins, including six glycoproteins currently named gE, gB, gH, gI, gC, and gL. Glycoproteins gE, gB, and gH are very abundant in infected cells and are also present in the virion envelope. Antibodies induced by three major glycoproteins can neutralize the virus. Specific humoral and cellular immunity, as well as cytokines such as interferons, play a significant role in suppressing and restoring VZV spread, with specific cellular immunity being particularly important.

[0194] Accordingly, in some embodiments, what is provided herein is a therapeutic nucleic acid encoding a viral peptide or protein derived from VZV. In some embodiments, the nucleic acid encodes a viral peptide or protein derived from VZV, the viral peptide or protein being one or more selected from (a) gE protein, (b) gB protein, (c) gH protein, (d) gI protein, (e) gC protein, (f) gL protein, (g) one immunogenic fragment from (a) to (f), and (h) one or more functional derivatives from (a) to (g).

[0195] Therefore, in some embodiments, the therapeutic nucleic acids of the present disclosure encode the VZV gE protein or an immunogenic fragment of the gE protein, or a functional derivative of the gE protein or its immunogenic fragment. Table 1 shows some exemplary VZV native antigen sequences. Table 1-1 Table 1-2 Table 1-3 Table 1-4 Table 1-5 Table 1-6 Table 1-7 Table 1-8 Table 1-9 Table 1-10 Table 1-11

[0196] In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mature gE protein of VZV, and the mature gE protein has the amino acid sequence of SEQ ID NO: 1. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mature gE protein of VZV, and the therapeutic nucleic acid includes the DNA coding sequence of SEQ ID NO: 2. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mature gE protein of VZV, and the therapeutic nucleic acid includes an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 2. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a full-length gE protein of VZV, and the full-length gE protein has the amino acid sequence of SEQ ID NO: 55. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a full-length gE protein of VZV, and the therapeutic nucleic acid includes the DNA coding sequence of SEQ ID NO: 56. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a full-length gE protein of VZV, and the therapeutic nucleic acid includes an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 56. In some embodiments, the RNA sequence is transcribed in vitro. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0197] In certain embodiments, the therapeutic nucleic acids of the present disclosure encode an immunogenic fragment of the VZV gE protein. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a functional derivative of the VZV gE protein. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode an immunogenic fragment of the VZV gE protein, the immunogenic fragment of the gE protein comprises a cleavage of at least one amino acid residue and up to 49 amino acid residues from the C-terminus compared to the mature gE protein. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode an immunogenic fragment of the gE protein of VZV, the immunogenic fragment of the gE protein comprising cleaving 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue compared to a mature gE protein. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes an immunogenic fragment of the gE protein of VZV, the immunogenic fragment of the gE protein comprising cleavage of 11–18 (e.g., 11, 12, 13, 14, 15, 16, 17, or 18) or 34–44 (e.g., 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44) amino acid residues from the C-terminus compared to the mature gE protein. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes an immunogenic fragment of the VZV gE protein, the immunogenic fragment comprising a cleavage of 14 or 37 amino acid residues from the C-terminus compared to the mature gE protein. In some embodiments, the RNA sequence is transcribed in vitro. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0198] In certain embodiments, the therapeutic nucleic acids of the Disclosure encode a variant of the VZV gE protein. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode a variant of the VZV gE protein, the variant comprising substitution Y569A. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode a variant of the VZV gE protein, the variant comprising substitution Y582G. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode a variant of the VZV gE protein, the variant comprising substitution Y569A and Y582G. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode a variant of the VZV gE protein, the variant comprising substitution S593A. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode a variant of the VZV gE protein, the variant comprising substitution S595A. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode a variant of the VZV gE protein, the variant comprising substitution T596A. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode variants of the VZV gE protein, the variants including substitution T598A. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode variants of the VZV gE protein, the variants including substitutions S593A, S595A, T596A, and T598A. In certain embodiments, the therapeutic nucleic acids of the Disclosure encode variants of the VZV gE protein, the variants including substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A. In such embodiments, amino acid positions are numbered based on the full-length gE protein. In some embodiments, the RNA sequence is transcribed in vitro. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0199] In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, the mutant fragment comprising cleavage of 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus compared to the mature gE protein. In such embodiments, the mutant fragment optionally comprises substitution Y569A. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, the mutant fragment comprising cleavage of 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, or 31 amino acid residues from the C-terminus compared to the mature gE protein. In such embodiments, the mutant fragment optionally comprises substitution Y582G, with or without substitution Y569A. In such embodiments, the mutant fragment comprises substitution Y569A and Y582G. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, the mutant fragment comprising a cleavage of 30 or 29 amino acid residues from the C-terminus compared to the mature gE protein. In such embodiments, the mutant fragment optionally comprises substitution S593A and is accompanied or absent with substitution Y569A and / or Y582G. In such embodiments, the mutant fragment comprises substitution Y569A, Y582G, and S593A. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, the mutant fragment comprising a cleavage of 28 amino acid residues from the C-terminus compared to the mature gE protein. In such embodiments, the mutant fragment optionally comprises substitution S595A and is accompanied or absent with substitution Y569A and / or Y582G and / or S593A. In such embodiments, the mutant fragment comprises substitution Y569A, Y582G, S593A, and S595A. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the VZV gE protein, the mutant fragment comprising a cleavage of 27 or 26 amino acid residues from the C-terminus compared to the mature gE protein. In such embodiments, the mutant fragment optionally comprises substitution T596A and is accompanied or absent by substitution Y569A and / or Y582G and / or S593A and / or S595A.In such embodiments, the mutant fragment includes substitutions Y569A, Y582G, S593A, S595A, and T596A. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the gE protein of VZV, the mutant fragment including cleavage of 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 amino acid residues, or 1 amino acid residue, compared to the mature gE protein. In such embodiments, the mutant fragment optionally includes substitution T598A and is accompanied or absent by substitutions Y569A and / or Y582G and / or S593A and / or S595A and / or T596A. In such embodiments, the mutant fragments include substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, the mutant fragment having the amino acid sequence of SEQ ID NO: 3. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid includes the DNA coding sequence of SEQ ID NO: 4 or 5. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid includes an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 4 or 5. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, the mutant fragment having the amino acid sequence of SEQ ID NO: 6. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid includes the DNA coding sequence of SEQ ID NO: 7. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 7. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the mutant fragment has the amino acid sequence of SEQ ID NO: 8. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid comprises the DNA coding sequence of SEQ ID NO: 9.In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid includes an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 9. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the mutant fragment has the amino acid sequence of SEQ ID NO: 10. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid includes the DNA coding sequence of SEQ ID NO: 11. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid includes an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 11. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the mutant fragment has the amino acid sequence of SEQ ID NO: 12. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid includes the DNA coding sequence of SEQ ID NO: 13. In certain embodiments, the therapeutic nucleic acid of the present disclosure encodes a mutant fragment of the VZV gE protein, and the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of Sequence ID No. 13. In some embodiments, the RNA sequence is transcribed in vitro. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0200] While not bound by theory, in some embodiments, the therapeutic nucleic acids of the present disclosure encode a fusion protein comprising a VZV gE protein or a fragment thereof fused to a trimerized peptide, so that the fusion protein can form a trimerized complex comprising three copies of the gE protein or a fragment thereof. In some embodiments, the gE protein or a fragment thereof is fused to the trimerized peptide via a peptide linker. Table 2 shows exemplary sequences of trimerized peptides and linker peptides, as well as fusion proteins, that can be used in connection with this disclosure. [Table 2]

[0201] In some embodiments, the therapeutic nucleic acid encodes a fusion protein comprising the gE protein of VZV or a functional derivative thereof fused to a trimerized peptide. In some embodiments, the fusion between the gE protein and the trimerized peptide is carried out via a peptide linker. In specific embodiments, the peptide linker comprises the amino acid sequence of SEQ ID NO: 14. In some embodiments, the trimerized peptide comprises the amino acid sequence of SEQ ID NO: 16.

[0202] In certain embodiments, the therapeutic nucleic acid encodes a fusion protein containing the gE protein of VZV fused to a trimerized peptide, and the nucleic acid contains a DNA coding sequence. In certain embodiments, the therapeutic nucleic acid encodes a fusion protein containing the gE protein of VZV fused to a trimerized peptide, and the nucleic acid contains an RNA sequence transcribed from the DNA coding sequence. In some embodiments, the RNA sequence is transcribed in vitro. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0203] While not bound by theory, fusion proteins containing a viral peptide or polypeptide fused to the Fc region of an immunoglobulin are intended to enhance the immunogenicity of the viral peptide or polypeptide. Therefore, in some embodiments, the therapeutic nucleic acid molecules of the present disclosure encode fusion proteins containing a VZV-derived viral peptide or protein fused to the Fc region of an immunoglobulin. In certain embodiments, the viral peptide or protein is one or more selected from (a) gE protein, (b) gB protein, (c) gH protein, (d) gI protein, (e) gC protein, (f) gL protein, (g) one immunogenic fragment from (a) to (f), and (h) one or more functional derivatives from (a) to (g). In certain embodiments, the immunoglobulin is human immunoglobulin (Ig). In certain embodiments, the immunoglobulin is human IgG, IgA, IgD, IgE, or IgM. In certain embodiments, the immunoglobulin is human IgG1, IgG2, IgG3, or IgG4. In some embodiments, immunoglobulin Fc is fused to the N-terminus of a viral peptide or polypeptide. In other embodiments, immunoglobulin Fc is fused to the C-terminus of a viral peptide or polypeptide.

[0204] While not bound by theory, signal peptides are intended to mediate the transport of the polypeptide to which they are fused to a specific location within a cell. Therefore, in some embodiments, the therapeutic nucleic acid molecules of the present disclosure encode a fusion protein comprising a viral peptide or protein fused to a signal peptide. In certain embodiments, the viral peptide or protein is one or more selected from (a) gE protein, (b) gB protein, (c) gH protein, (d) gI protein, (e) gC protein, (f) gL protein, (g) one immunogenic fragment from (a) to (f), and (h) one or more functional derivatives from (a) to (g). In some embodiments, the signal peptide is fused to the N-terminus of the viral peptide or polypeptide. In other embodiments, the signal peptide is fused to the C-terminus of the viral peptide or polypeptide. Table 3 shows exemplary sequences of signal peptides that can be used in connection with the present disclosure, and exemplary VZV antigen sequences containing the signal peptide. [Table 3-1] [Table 3-2]

[0205] In certain embodiments, the signal peptide is encoded by a VZV gene from which a viral peptide or polypeptide is derived. In certain embodiments, the signal peptide encoded by a VZV gene is fused to a viral peptide or polypeptide encoded by a different VZV gene. In other embodiments, the signal peptide encoded by a VZV gene is fused to a viral peptide or polypeptide encoded by the same VZV gene. For example, in some embodiments, a signal peptide having the amino acid sequence MGTVNKPVVGVLMGFGIITGTLRITNPVRA (SEQ ID NO: 18) is fused to a viral peptide or polypeptide encoded by a nucleic acid molecule of the present disclosure. In various embodiments, the viral peptide or protein is one or more selected from (a) E protein, (b) B protein, (c) H protein, (d) I protein, (e) C protein, (f) L protein, (g) one immunogenic fragment from (a) to (f), and (h) one or more functional derivatives from (a) to (g).

[0206] In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a VZV gE protein or fragment that does not have a native signal peptide. In certain embodiments, the encoded gE protein or fragment contains a signal peptide having the amino acid sequence of SEQ ID NO: 23 or 27. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a VZV gE protein or fragment having a signal peptide, and the therapeutic nucleic acid contains the DNA coding sequence of SEQ ID NO: 24, 25, 26, or 28. In certain embodiments, the therapeutic nucleic acid of the Disclosure encodes a VZV gE protein or fragment having a signal peptide, and the therapeutic nucleic acid contains an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 24, 25, 26, or 28. In some embodiments, the RNA sequence is transcribed in vitro. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0207] In other embodiments, the signal peptide is encoded by an exogenous gene sequence not present in the VZV from which the viral peptide or polypeptide is derived. In some embodiments, the heterologous signal peptide replaces a homologous signal peptide in a fusion protein encoded by the nucleic acid molecule of the Disclosure. In specific embodiments, the signal peptide is encoded by a mammalian gene. In specific embodiments, the signal peptide is encoded by a human immunoglobulin gene. In specific embodiments, the signal peptide is encoded by a human IgE gene. For example, in some embodiments, a signal peptide having the amino acid sequence MDWTWILFLVAAATRVHS (SEQ ID NO: 23) is fused to a viral peptide or polypeptide encoded by the nucleic acid molecule of the Disclosure. In various embodiments, the viral peptide or protein is one or more selected from (a) E protein, (b) B protein, (c) H protein, (d) I protein, (e) C protein, (f) L protein, (g) one immunogenic fragment from (a) to (f), and (h) one or more functional derivatives from (a) to (g).

[0208] In some embodiments, what is provided herein is a nucleic acid encoding a fragment of mature gE as described in Section 5.3. In some embodiments, what is provided herein is a nucleic acid encoding a fusion protein as described in Section 5.3. In some embodiments, what is provided herein is a nucleic acid encoding a variant of full-length VZV gE as described in Section 5.3. In some embodiments, what is provided herein is a nucleic acid encoding a protein as described in Section 5.3. In some embodiments, what is provided herein is a nucleic acid comprising the nucleotide sequence described in SEQ ID NOs: 7, 9, 11, 13, 4, or 5. In some embodiments, what is provided herein is a nucleic acid comprising a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the nucleotide sequence described in SEQ ID NOs: 7, 9, 11, 13, 4, or 5. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA and comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-psoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages described in section 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure described in section 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-untranslated region (5'-UTR) described in section 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3'-untranslated region (3'-UTR) described in section 5.4.3.In some embodiments, the nucleic acid further comprises, for example, a 5' untranslated region (5'-UTR) as described in Section 5.4.3 and, for example, a 3' untranslated region (3'-UTR) as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises a nucleotide sequence as described in any one of SEQ ID NOs. 29-38. In some embodiments, the 3'-UTR comprises a nucleotide sequence as described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence as described in any one of SEQ ID NOs. 29-38 and the 3'-UTR comprises a nucleotide sequence as described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or polyadenylation signal (for example, as described in Section 5.4.4).

[0209] In some embodiments, provided herein is a nucleic acid encoding a protein or fusion protein as described in Section 5.3, comprising a variant or fragment of mature gE and a human IgE signal peptide, wherein the nucleotide sequence encoding the IgE signal peptide comprises the nucleotide sequence described in SEQ ID NO: 24, 25, or 26. In some embodiments, provided herein is a nucleic acid encoding a protein or fusion protein as described in Section 5.3, comprising a variant or fragment of mature gE and a human IgE signal peptide, wherein the nucleotide sequence encoding the IgE signal peptide comprises a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 24, 25, or 26. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA and comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-psoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages described in section 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure described in section 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-untranslated region (5'-UTR) described in section 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3'-untranslated region (3'-UTR) described in section 5.4.3. In some embodiments, the nucleic acid further includes, for example, a 5' untranslated region (5'-UTR) as described in Section 5.4.3 and, for example, a 3' untranslated region (3'-UTR) as described in Section 5.4.3.In some embodiments, the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38, and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal (e.g., those described in Section 5.4.4).

[0210] In some embodiments, what is provided herein is a nucleic acid encoding a protein as described in Section 5.3, comprising a variant of mature gE and a signal peptide, wherein the nucleotide sequence encoding the signal peptide comprises the nucleotide sequence described in SEQ ID NOs: 19, 20, 21, or 22. In some embodiments, what is provided herein is a nucleic acid encoding a protein as described in Section 5.3, comprising a variant of mature gE and a signal peptide, wherein the nucleotide sequence encoding the signal peptide is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NOs: 19, 20, 21, or 22. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in Sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA and comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methylpsoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages as described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-untranslated region (5'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 3'-untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further includes a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3).In some embodiments, the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38, and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal (e.g., those described in Section 5.4.4).

[0211] In some embodiments, what is provided herein is a nucleic acid encoding a protein or fusion protein as described in Section 5.3, comprising a variant or fragment of mature gE and a human tPA signal peptide, wherein the nucleotide sequence encoding the human tPA signal peptide comprises the nucleotide sequence described in SEQ ID NO: 28. In some embodiments, what is provided herein is a nucleic acid encoding a protein or fusion protein as described in Section 5.3, comprising a variant or fragment of mature gE and a human tPA signal peptide, wherein the nucleotide sequence encoding the human tPA signal peptide comprises a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 28. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs as described in Sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA and comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methylpsoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages as described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-untranslated region (5'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 3'-untranslated region (3'-UTR) (e.g., as described in Section 5.4.3).In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38 and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or polyadenylation signal (e.g., as described in Section 5.4.4).

[0212] In some embodiments, what is provided herein is a nucleic acid comprising the nucleotide sequence described in SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, what is provided herein is a nucleic acid comprising the nucleotide sequence described in SEQ ID NOs: 63. In some embodiments, what is provided herein is a nucleic acid consisting of, essentially therefor, or comprising the nucleotide sequence described in SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, what is provided herein is a nucleic acid consisting of, essentially therefor, or comprising the nucleotide sequence described in SEQ ID NOs: 63. In some embodiments, what is provided herein is a nucleic acid consisting of, or containing, a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identity with the nucleotide sequence described in SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, what is provided herein is a nucleic acid consisting of, or containing, a nucleotide sequence having at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identity with the nucleotide sequence described in SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, what is provided herein are nucleic acids consisting of, essentially, or containing nucleotide sequences having at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identity with the nucleotide sequences described in SEQ ID NOs. 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5.In some embodiments, what is provided herein is a nucleic acid consisting of, essentially, or containing a nucleotide sequence having at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequences described in SEQ ID NOs. 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-psoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages as described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the 5'-UTR comprises a nucleotide sequence as described in any one of SEQ ID NOs. 29 to 38. In some embodiments, the 3'-UTR comprises the nucleotide sequence described in any one of sequence numbers 39-46.In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38, and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal (e.g., those described in Section 5.4.4).

[0213] In some embodiments, the nucleic acid provided herein encodes the amino acid sequence described in SEQ ID NO: 6. In some embodiments, the nucleic acid provided herein comprises the nucleotide sequence described in SEQ ID NO: 7. In some embodiments, the nucleic acid provided herein encodes the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the nucleic acid comprises the nucleotide sequence described in SEQ ID NO: 9. In some embodiments, the nucleic acid provided herein encodes the amino acid sequence described in SEQ ID NO: 10. In some embodiments, the nucleic acid comprises the nucleotide sequence described in SEQ ID NO: 11. In some embodiments, the nucleic acid provided herein encodes the amino acid sequence described in SEQ ID NO: 12. In some embodiments, the nucleic acid comprises the nucleotide sequence described in SEQ ID NO: 13. In some embodiments, the nucleic acid provided herein encodes the amino acid sequence described in SEQ ID NO: 3. In some embodiments, the nucleic acid comprises the nucleotide sequence described in SEQ ID NO: 4 or 5. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA and comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methylpsoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages as described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-untranslated region (5'-UTR) (e.g., as described in Section 5.4.3).In some embodiments, the nucleic acid further comprises a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38 and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or polyadenylation signal (e.g., as described in Section 5.4.4).

[0214] In some embodiments, what is provided herein is a nucleic acid encoding the amino acid sequence of SEQ ID NO: 59. In some embodiments, the nucleic acid comprises the nucleotide sequence described in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid comprises the nucleotide sequence described in SEQ ID NO: 63. In some embodiments, the nucleic acid consists of the nucleotide sequence described in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid consists of the nucleotide sequence described in SEQ ID NO: 63. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identical to the nucleotide sequence described in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identical to the nucleotide sequence described in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identical to the nucleotide sequence described in SEQ ID NOs. 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the nucleotide sequence described in SEQ ID NOs. 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-psoiduridine, and 5-methylcytosine.In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages as described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the 5'-UTR comprises a nucleotide sequence as described in any one of SEQ ID NOs. 29 to 38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38, and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal (e.g., those described in Section 5.4.4).

[0215] In some embodiments, what is provided herein is a nucleic acid comprising the nucleotide sequence described in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, what is provided herein is a nucleic acid consisting of the nucleotide sequence described in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, what is provided herein is a nucleic acid consisting of, or comprising, a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identity with the nucleotide sequence described in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, what is provided herein is a nucleic acid consisting of, or comprising, a nucleotide sequence having at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identity with the nucleotide sequence described in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the herein provides nucleic acids consisting of, or comprising, nucleotide sequences having at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identity with the nucleotide sequences described in SEQ ID NOs. 51, 60, 61, 62, 63, or 64. In some embodiments, the herein provides nucleic acids consisting of, or comprising, nucleotide sequences having at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the nucleotide sequences described in SEQ ID NOs. 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8).In some embodiments, the nucleic acid is mRNA and comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methylpsoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages as described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-untranslated region (5'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 3'-untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38 and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or polyadenylation signal (e.g., as described in Section 5.4.4).

[0216] In some embodiments, what is provided herein is a nucleic acid comprising the nucleotide sequence described in Sequence ID No. 63. In some embodiments, what is provided herein is a nucleic acid consisting of the nucleotide sequence described in Sequence ID No. 63. In some embodiments, what is provided herein is a nucleic acid consisting of, or comprising, a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identity with the nucleotide sequence described in Sequence ID No. 63. In some embodiments, what is provided herein is a nucleic acid consisting of, or comprising, a nucleotide sequence having at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identity with the nucleotide sequence described in Sequence ID No. 63. In some embodiments, what is provided herein is a nucleic acid consisting of, or comprising, a nucleotide sequence having at least 90%, at least 91%, at least 92%, at least 93%, or at least 94% identity with the nucleotide sequence described in Sequence ID No. 63. In some embodiments, what is provided herein is a nucleic acid consisting of, or containing, a nucleotide sequence having at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence described in SEQ ID NO: 63. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA and comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-psoiduridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages described in section 5.4.9).In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38, and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal (e.g., those described in Section 5.4.4).

[0217] In some embodiments, what is provided herein is a nucleic acid comprising the nucleotide sequence described in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, what is provided herein is a nucleic acid consisting of the nucleotide sequence described in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, what is provided herein is a nucleic acid consisting of, or comprising, a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, or at least 84% identity with the nucleotide sequence described in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, what is provided herein is a nucleic acid consisting of, or comprising, a nucleotide sequence having at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% identity with the nucleotide sequence described in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, what is provided herein is a nucleic acid consisting of, or containing, at least 90%, at least 91%, at least 92%, or at least 93% of the nucleotide sequence 49, 50, 52, 53, or 54. In some embodiments, what is provided herein is a nucleic acid consisting of, or containing, a nucleotide sequence having at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence described in SEQ ID NOs. 49, 50, 52, 53, or 54. In some embodiments, the nucleic acid does not exist in nature. In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methylpsoiduridine, and 5-methylcytosine.In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages as described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the 5'-UTR comprises a nucleotide sequence as described in any one of SEQ ID NOs. 29 to 38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38, and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal (e.g., those described in Section 5.4.4).

[0218] In some embodiments, what is provided herein is the nucleic acid listed in Table 1, or the mRNA of the nucleic acid listed in Table 1. In some embodiments, what is provided herein is the nucleic acid listed in Table 1 other than SEQ ID NOs. 2 and 56, or the mRNA of the nucleic acid listed in Table 1 other than SEQ ID NOs. 2 and 56. In some embodiments, what is provided herein is the nucleic acid containing the VZV gE mutain coding sequence listed in Table 1, or the mRNA of the nucleic acid of the VZV gE mutain coding sequence listed in Table 1. In some embodiments, what is provided herein is the nucleic acid consisting of the VZV gE mutain coding sequence listed in Table 1, or the mRNA of the nucleic acid of the VZV gE mutain coding sequence listed in Table 1. In some embodiments, what is provided herein is the nucleic acid containing the VZV gE mutain-1 coding sequence listed in Table 1, or the mRNA of the nucleic acid of the VZV gE mutain-1 coding sequence listed in Table 1. In some embodiments, what is provided herein is the nucleic acid containing the VZV gE mutain-2 coding sequence listed in Table 1, or the mRNA of the nucleic acid of the VZV gE mutain-2 coding sequence listed in Table 1. In some embodiments, what is provided herein is a nucleic acid containing the VZV gE mutain-3 coding sequence listed in Table 1, or mRNA of the nucleic acid of the VZV gE mutain-3 coding sequence listed in Table 1. In some embodiments, what is provided herein is a nucleic acid containing the VZV gE mutain-4 coding sequence listed in Table 1, or mRNA of the nucleic acid of the VZV gE mutain-4 coding sequence listed in Table 1. In some embodiments, what is provided herein is a nucleic acid containing the VZV gE mutain-5 coding sequence listed in Table 1, or mRNA of the nucleic acid of the VZV gE mutain-5 coding sequence listed in Table 1. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs (e.g., one or more functional nucleotide analogs described in sections 5.4.6, 5.4.7, and / or 5.4.8). In some embodiments, the nucleic acid is mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-psoiduridine, and 5-methylcytosine.In some embodiments, the nucleic acid comprises one or more modified nucleoside linkages (e.g., one or more modified nucleoside linkages as described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5'-cap structure (e.g., as described in Section 5.4.2). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR) (e.g., as described in Section 5.4.3) and a 3' untranslated region (3'-UTR) (e.g., as described in Section 5.4.3). In some embodiments, the 5'-UTR comprises a nucleotide sequence as described in any one of SEQ ID NOs. 29 to 38. In some embodiments, the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, where the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29-38, and the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39-46. In some embodiments, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal (e.g., those described in Section 5.4.4).

[0219] In specific embodiments, the coding nucleotide sequences of nucleic acids described herein (e.g., nucleic acids not found in nature) are codon-optimized for expression in the target cell, and the target is optionally a non-human mammal or human.

[0220] In specific embodiments, the nucleic acids described herein (e.g., nucleic acids not found in nature) are mRNA, and thymine (e.g., in the corresponding DNA sequence) is replaced within the nucleic acid by uracil or a functional analogue.

[0221] In specific embodiments, the nucleic acids provided herein are those described in Section 6 below. In specific embodiments, the codon-optimized nucleic acids provided herein are those described in Section 6 below. In specific embodiments, the nucleic acids provided herein are those not found in nature, those described in Section 6 below. In some embodiments, the nucleic acids provided herein encode variants of VZV gE other than the control described in Section 6 below.

[0222] 5.4.2 5' Cap Structure While not bound by theory, the 5' cap structure of polynucleotides is intended to be involved in nuclear export and increased polynucleotide stability, and to bind to mRNA cap-binding proteins (CBPs), which are responsible for intracellular polynucleotide stability and translational capacity by associating with poly(A)-binding proteins to form mature circular mRNA species. The 5' cap structure further assists in the removal of 5' proximal introns during mRNA splicing. Therefore, in some embodiments, the nucleic acid molecules of this disclosure include a 5' cap structure.

[0223] Nucleic acid molecules can be capped at the 5' end by the cell's endogenous transcription mechanism, creating a 5'-ppp-5'-triphosphate linkage between the terminal guanosine cap residue of the polynucleotide and the 5'-terminal transcription sense nucleotide. This 5' guanylate cap can then be methylated to produce an N7-methyl-guanylate residue. The ribose sugars of the 5' terminal and / or anteterminal transcription nucleotides of the polynucleotide can also be optionally 2'-O-methylated. 5' decapping and cleavage of the guanylate cap structure via hydrolysis can target nucleic acid molecules such as mRNA molecules for degradation.

[0224] In some embodiments, the nucleic acid molecules of this disclosure include one or more modifications to the native 5' cap structure produced by endogenous processes. While not limited to theory, modifications on the 5' cap may increase the stability of the polynucleotide, increase the half-life of the polynucleotide, and increase the translation efficiency of the polynucleotide.

[0225] Exemplary modifications to the natural 5' cap structure include the creation of a non-hydrolyzable cap structure that prevents detachment and thus increases the polynucleotide half-life. In some embodiments, hydrolysis of the cap structure requires cleavage of the 5'-ppp-5' phosphorodiester linkage; therefore, in some embodiments, modified nucleotides may be used during the capping reaction. For example, in some embodiments, a vaccinia capping enzyme from New England Biolabs (Ipswich, Mass.) may be used with α-thio-guanosine nucleotides, according to the manufacturer's instructions, to create the phosphorothioate linkage at the 5'-ppp-5' cap. Additional modified guanosine nucleotides, such as α-methyl-phosphonate and seleno-phosphate nucleotides, may be used.

[0226] Additional exemplary modifications to the natural 5' cap structure also include modifications at the 2' and / or 3' positions of the capped guanosine triphosphate (GTP), replacement of the methylene moiety (CH2) of the sugar ring oxygen (which formed the carbocyclic ring), modifications at the triphosphate crosslinking portion of the cap structure, or modifications at the nucleic acid base (G) portion.

[0227] Additional exemplary modifications to the natural 5'-cap structure include, but are not limited to, 2'-O-methylation of the ribose sugar of the 5'-terminus and / or 5'-pre-terminus nucleotide of a polynucleotide (as described above) on the 2'-hydroxyl group of the sugar. Multiple different 5'-cap structures can be used to generate the 5'-cap of a polynucleotide, such as an mRNA molecule. Additional exemplary 5'-cap structures that can be used in connection with this disclosure are further described in International Patent Publications WO2008 / 127688, WO2008 / 016473, and WO2011 / 015347, the entire contents of each of which are incorporated herein by reference.

[0228] In various embodiments, the 5'-terminal cap may include cap analogues. Cap analogues, also referred to herein as synthetic cap analogues, chemical caps, chemical cap analogues, or structural or functional cap analogues, retain cap function while differing in their chemical structure from the natural (i.e., endogenous, wild-type, or physical) 5' cap. Cap analogues may be synthesized chemically (i.e., non-enzymatically) or enzymatically and / or linked to polynucleotides.

[0229] For example, an anti-reverse cap analog (ARCA) cap contains two guanosines linked by a 5'-5'-triphosphate group, one of which contains an N7-methyl group and a 3'-O-methyl group (i.e., N7,3'-O-dimethyl-guanosine-5'-triphosphate-5'-guanosine, m7G-3'mppp-G, which can be equivalently written as 3'O-Me-m7G(5')ppp(5')G). The 3'-O atom of the other unmodified guanosine is then linked to the 5' terminal nucleotide of the capped polynucleotide (e.g., mRNA). N7-methylated and 3'-O-methylated guanosines provide terminal portions of the capped polynucleotide (e.g., mRNA). Another exemplary cap structure is mCAP, which is similar to ARCA but has a 2'-O-methyl group on guanosine (i.e., N7,2'-O-dimethyl-guanosine-5'-triphosphate-5'-guanosine, m7Gm-ppp-G).

[0230] In some embodiments, the cap analog may be a dinucleotide cap analog. In non-limiting examples, the dinucleotide cap analog may be modified at different phosphate positions with a boranophosphate group or a phosphoroselenoate group, such as the dinucleotide cap analog described in U.S. Patent No. 8,519,110, the entire scope of which is incorporated herein by reference.

[0231] In some embodiments, the cap analogues may be N7-(4-chlorophenoxyethyl)-substituted dinucleotide cap analogues known in the art and / or described herein. Non-limiting examples of N7-(4-chlorophenoxyethyl)-substituted dinucleotide cap analogues include N7-(4-chlorophenoxyethyl)-G(5')ppp(5')G and N7-(4-chlorophenoxyethyl)-m3'-OG(5')ppp(5')G cap analogues (see, for example, Kore et al. Bioorganic & Medicinal Chemistry 2013 21:4570-4574, the full contents of which are incorporated herein by reference). In other embodiments, a useful cap analogue in relation to the nucleic acid molecules of this disclosure is a 4-chloro / bromophenoxyethyl analogue.

[0232] In various embodiments, the cap analog may include guanosine analogs. Useful guanosine analogs include, but are not limited to, inosine, N1-methyl-guanosine, 2'-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

[0233] While not bound by theory, cap analogs allow for the simultaneous capping of polynucleotides, but in in vitro transcription reactions, it is intended that up to 20% of the transcript remains uncapped. This, like the structural differences between cap analogs and the natural 5' cap structure of polynucleotides produced by the cell's endogenous transcription mechanism, can lead to reduced translational capacity and decreased cellular stability.

[0234] Accordingly, in some embodiments, nucleic acid molecules of the present disclosure may be post-transcriptionally capped using an enzyme to produce a more authentic 5' cap structure. As used herein, the term “more authentic” refers to a feature that more closely reflects or mimics an endogenous or wild-type feature, either structurally or functionally. That is, a “more authentic” feature better represents an endogenous wild-type, natural, or physiological cellular function and / or structure, or is superior in one or more respects to the corresponding endogenous wild-type, natural, or physiological feature, compared to a synthetic feature or analogue of the prior art. Non-limiting examples of more authentic 5' cap structures useful in relation to nucleic acid molecules of the present disclosure include, among other things, those having enhanced cap-binding protein binding, increased half-life, reduced sensitivity to 5' endonucleases, and / or reduced 5' decapping, compared to synthetic 5' cap structures (or wild-type, natural, or physiological 5' cap structures) known in the art. For example, in some embodiments, recombinant vaccinia virus capping enzyme and recombinant 2'-O-methyltransferase enzyme can create a canonical 5'-5'-triphosphate bond between the 5' terminal nucleotides of a polynucleotide and a guanosine cap nucleotide, wherein the cap guanosine includes N7 methylation and the 5' terminal nucleotide of the polynucleotide includes 2'-O-methylation. Such a structure is referred to as a cap 1 structure. This cap provides, for example, higher translational qualification, cellular stability, and reduced activation of cellular inflammatory cytokines compared to other 5' cap analog structures known in the art. Other exemplary cap structures include 7mG(5')ppp(5')N,pN2p (cap 0), 7mG(5')ppp(5')NlmpNp (cap 1), 7mG(5')-ppp(5')NlmpN2mp (cap 2), and m(7)Gpppm(3)(6,6,2')Apm(2')Apm(2')Cpm(2)(3,2')Up (cap 4). In some embodiments, the cap structure is m 7 Includes GpppAmpU.

[0235] While not bound by theory, the nucleic acid molecules of this disclosure can be capped after transcription, and since this process is more efficient, it is intended that nearly 100% of nucleic acid molecules can be capped.

[0236] In some embodiments provided herein, all thymine (T) is substituted with uracil (U) or N1-methylpsoiduridine, and / or the first nucleotide G is m 7 Except for being substituted with GpppAmpU, (1) the nucleotide described in SEQ ID NO: 63, or (2) a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 63, is a nucleic acid that is essentially derived from or contains the same. In some embodiments, provided herein are nucleic acids in which all thymine (T) is substituted with uracil (U) or N1-methylpsoiduridine, and / or the first nucleotide G is m 7 Nucleic acids comprising, or containing, (1) the nucleotide sequence described in SEQ ID NO: 63, or (2) a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 63, except that it is substituted with GpppAmpU. In some embodiments, provided herein are nucleic acids comprising, or containing, a nucleotide sequence comprising (1) the nucleotide sequence described in SEQ ID NO: 63, except that it is substituted with GpppAmpU, or (2) a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 63 7 A nucleic acid consisting of, essentially, or containing the nucleotide sequence described in Sequence ID No. 63, except that it is substituted with GpppAmpU. In some embodiments, provided herein is a nucleic acid in which all thymine (T) is substituted with N1-methylpsoiduridine, and the first nucleotide G is m 7 A nucleic acid consisting of, essentially, or containing the nucleotide sequence described in SEQ ID NO: 63, except that it is substituted with GpppAmpU.

[0237] 5.4.3 Untranslated Area (UTR) In some embodiments, the nucleic acid molecules of this disclosure include one or more untranslated regions (UTRs). In some embodiments, the UTRs are located upstream of the coding region in the nucleic acid molecule and are referred to as 5'-UTRs. In some embodiments, the UTRs are located downstream of the coding region in the nucleic acid molecule and are referred to as 3'-UTRs. The sequences of the UTRs may be homologous or heterologous to the sequences of the coding region found in the nucleic acid molecule. Multiple UTRs may be included in the nucleic acid molecule, and may have the same or different sequences and / or be of genetic origin. According to this disclosure, any portion (including none) of the UTRs in the nucleic acid molecule may be codon-optimized, and each may independently contain one or more different structural or chemical modifications before and / or after codon optimization.

[0238] In some embodiments, the nucleic acid molecule (e.g., mRNA) of the Disclosure comprises a UTR and coding regions that are homologous to each other. In other embodiments, the nucleic acid molecule (e.g., mRNA) of the Disclosure comprises a UTR and coding regions that are heterogeneous to each other. In some embodiments, to monitor the activity of the UTR sequence, a nucleic acid molecule comprising a UTR and a coding sequence of a detectable probe can be administered in vitro (e.g., in a cell or tissue culture) or in vivo (e.g., in a subject), and the effect of the UTR sequence (e.g., regulation of expression level, cellular localization of the encoded product, or half-life of the encoded product) can be measured using methods known in the art.

[0239] In some embodiments, the UTR of a nucleic acid molecule (e.g., mRNA) of the Disclosure includes at least one translational enhancer element (TEE) that functions to increase the amount of polypeptide or protein produced from the nucleic acid molecule. In some embodiments, the TEE is located at the 5'-UTR of the nucleic acid molecule. In other embodiments, the TEE is located at the 3'-UTR of the nucleic acid molecule. In yet another embodiment, at least two TEEs are located at the 5'-UTR and 3'-UTR of the nucleic acid molecule, respectively. In some embodiments, a nucleic acid molecule (e.g., mRNA) of the Disclosure may contain one or more copies of a TEE sequence, or two or more different TEE sequences. In some embodiments, the different TEE sequences present in the nucleic acid molecule of the Disclosure may be homologous or heterogeneous with respect to one another.

[0240] Various TEE sequences known in the art may be used in connection with this disclosure. For example, in some embodiments, the TEE may be an internal ribosome entry site (IRES), an HCV-IRES, or an IRES element. Chappell et al. Proc.Natl.Acad.Sci.USA 101:9590-9594,2004; Zhou et al. Proc.Natl.Acad.Sci.102:6273-6278,2005. Additional internal ribosome entry sites (IRESs) that can be used in connection with this disclosure include, but are not limited to, those described in U.S. Patent No. 7,468,275, U.S. Patent Publication No. 2007 / 0048776, and U.S. Patent Publication No. 2011 / 0124100, and International Patent Publication No. WO2007 / 025008 and International Patent Publication No. WO2001 / 055369, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the TEE may be those described in Supplemental Tables 1 and 2 of Wellensiek et al. Genome-wide profiling of human cap-independent translation-enhancing elements, Nature Methods, 2013 Aug;10(8): 747-750, the contents of which are incorporated by reference in their entirety.

[0241] Additional exemplary TEEs that may be used in connection with this disclosure include U.S. Patent Nos. 6,310,197, 6,849,405, 7,456,273, 7,183,395, U.S. Patent Publication No. 2009 / 0226470, U.S. Patent Publication No. 2013 / 0177581, U.S. Patent Publication No. 2007 / 0048776, U.S. Patent Publication No. 2011 / 0124100, and U.S. Patent Publication No. 2009 / 0093049. Examples include, but are not limited to, the TEE sequences disclosed in International Patent Publication WO2009 / 075886, International Patent Publication WO2012 / 009644, and International Patent Publication WO1999 / 024595, International Patent Publication WO2007 / 025008, International Patent Publication WO2001 / 055371, European Patent No. 2610341, and European Patent No. 2610340, the contents of which are incorporated herein by reference in their entirety.

[0242] In various embodiments, a nucleic acid molecule (e.g., mRNA) of the Disclosure includes at least one UTR containing at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, or more than 60 TEE sequences. In some embodiments, the TEE sequences in the UTR of the nucleic acid molecule are copies of the same TEE sequence. In other embodiments, at least two TEE sequences in the UTR of the nucleic acid molecule are different TEE sequences. In some embodiments, multiple different TEE sequences are arranged in one or more repeating patterns in the UTR region of the nucleic acid molecule. For illustrative purposes only, repeating patterns may be, for example, ABABAB, AABBAABBAABB, ABCABCABC, and in these exemplary patterns, each capital letter (A, B, or C) represents a different TEE sequence. In some embodiments, at least two TEE sequences are consecutive to each other (i.e., there are no spacer sequences in between) in the UTR of the nucleic acid molecule. In other embodiments, at least two TEE sequences are separated by a spacer sequence. In some embodiments, the UTR may include a TEE sequence spacer sequence module that is repeated at least once, at least twice, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or more than nine times in the UTR. In any of the embodiments described in this paragraph, the UTR may be the 5'-UTR, 3'-UTR, or both the 5'-UTR and 3'-UTR of the nucleic acid molecule.

[0243] In some embodiments, the UTR of a nucleic acid molecule (e.g., mRNA) of the Disclosure comprises at least one translational repressor element that functions to reduce the amount of polypeptide or protein produced from the nucleic acid molecule. In some embodiments, the UTR of a nucleic acid molecule comprises one or more miR sequences or fragments thereof (e.g., miR seed sequences) recognized by one or more microRNAs. In some embodiments, the UTR of a nucleic acid molecule comprises one or more stem-loop structures that downregulate the translational activity of the nucleic acid molecule. Other mechanisms for repressing translational activity associated with nucleic acid molecules are known in the Art. In any of the embodiments described in this paragraph, the UTR may be a 5'-UTR, a 3'-UTR, or both a 5'-UTR and a 3'-UTR of the nucleic acid molecule. Table 4 shows exemplary 5'-UTR and 3'-UTR sequences that can be used in connection with the Disclosure. [Table 4-1] [Table 4-2]

[0244] In specific embodiments, the nucleic acid molecule of the Disclosure comprises a 5'-UTR selected from any one of SEQ ID NOs: 29-38. In specific embodiments, the nucleic acid molecule of the Disclosure comprises a 3'-UTR selected from any one of SEQ ID NOs: 39-46. In specific embodiments, the nucleic acid molecule of the Disclosure comprises a 5'-UTR selected from any one of SEQ ID NOs: 29-38 and a 3'-UTR selected from any one of SEQ ID NOs: 39-46. In any of the embodiments described in this paragraph, the nucleic acid molecule may further comprise a coding region having a sequence described herein, for example, any of the DNA coding sequences in Tables 1-4, or an equivalent RNA sequence thereof. In certain embodiments, the nucleic acid molecule described in this paragraph may be an RNA molecule transcribed in vitro. [Table 5-1] [Table 5-2] Table 5-3 Table 5-4 Table 5-5 Table 5-6 Table 5-7 Table 5-8 Table 5-9 Table 5-10 Table 5-11 Table 5-12 Table 5-13 Table 5-14 Table 5-15 Table 5-16 Table 5-17 Table 5-18 Table 5-19 [Table 5-20]

[0245] 5.4.4 Polyadenylated (Poly-A) Region During natural RNA processing, long adenosine nucleotides (poly-A regions) are typically added to messenger RNA (mRNA) molecules, increasing their stability. Immediately after transcription, the 3' end of the transcript is cleaved to release 3'-hydroxyl. Poly-A polymerase then adds the adenosine nucleotide chain to the RNA. This process, called polyadenylation, adds poly-A regions between 100 and 250 residues in length. While not theoretically bound, poly-A regions are thought to confer various advantages to the nucleic acid molecules of this disclosure.

[0246] Accordingly, in some embodiments, the nucleic acid molecule (e.g., mRNA) of the Disclosure includes a polyadenylation signal. In some embodiments, the nucleic acid molecule (e.g., mRNA) of the Disclosure includes one or more polyadenylation (poly-A) regions. In some embodiments, the poly-A region consists entirely of an adenine nucleotide or a functional analog thereof. In some embodiments, the nucleic acid molecule includes at least one poly-A region at its 3' end. In some embodiments, the nucleic acid molecule includes at least one poly-A region at its 5' end. In some embodiments, the nucleic acid molecule includes at least one poly-A region at its 5' end and at least one poly-A region at its 3' end.

[0247] According to this disclosure, the length of the polyA region can vary in different embodiments. In particular, in some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 30 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 35 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 40 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 45 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 50 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 55 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 60 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 65 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 70 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of this disclosure is at least 75 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 80 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 85 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 90 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 95 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 100 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 110 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 120 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 130 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 140 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 150 nucleotides long.In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 160 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 170 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 180 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 190 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 200 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 225 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 250 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 275 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 300 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 350 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 400 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 450 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 500 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 600 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 700 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 800 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 900 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1000 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1100 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1200 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1300 nucleotides long.In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1400 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1500 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1600 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1700 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1800 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 1900 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 2000 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 2250 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 2500 nucleotides long. In some embodiments, the polyA region of the nucleic acid molecule of the disclosure is at least 2750 nucleotides long. In some embodiments, the poly(A) region of the nucleic acid molecule of the present disclosure is at least 3000 nucleotides long.

[0248] In some embodiments, the length of the polyA region in a nucleic acid molecule may be selected based on the total length of the nucleic acid molecule or a portion thereof (e.g., the length of the coding region or the length of the open reading frame of the nucleic acid molecule). For example, in some embodiments, the polyA region occupies about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more of the total length of the nucleic acid molecule containing the polyA region.

[0249] While not theoretically bound, certain RNA-binding proteins are thought to be able to bind to the poly(A) region located at the 3' end of the mRNA molecule. These poly(A)-binding proteins (PABPs) can regulate mRNA expression, for example, by interacting with the translation initiation mechanism in cells and / or protecting the 3' poly(A) tail from degradation. Therefore, in some embodiments, the nucleic acid molecule (e.g., mRNA) of this disclosure contains at least one binding site for a poly(A)-binding protein (PABP). In other embodiments, the nucleic acid molecule is coupled to or complexed with a PABP before being loaded onto a delivery vehicle (e.g., lipid nanoparticles).

[0250] In some embodiments, the nucleic acid molecule (e.g., mRNA) of this disclosure comprises a polyAG quartet. The G quartet is a cyclic hydrogen-bonded array of four guanosine nucleotides that can be formed by a G-rich sequence in both DNA and RNA. In this embodiment, the G quartet is incorporated at the end of a polyA region. The resulting polynucleotide (e.g., mRNA) can be assayed for stability, protein production, and other parameters, including half-life, at various time points. It has been found that the polyAG quartet structure yields protein production equal to at least 75% of the protein production seen using only a 120-nucleotide polyA region.

[0251] In some embodiments, the nucleic acid molecules of this disclosure (e.g., mRNA) may include a poly-A region and may be stabilized by the addition of a 3'-stabilizing region. In some embodiments, 3'-stabilizing regions that can be used to stabilize nucleic acid molecules (e.g., mRNA) including a poly-A or poly-AG quartet structure are such as those described in International Patent Publication WO2013 / 103659, the contents of which are incorporated herein by reference in their entirety.

[0252] In other embodiments, 3'-stabilizing regions that can be used in relation to nucleic acid molecules of this disclosure include 3'-deoxyadenosine (cordycepin), 3'-deoxyuridine, 3'-deoxycytosine, 3'-deoxyguanosine, 3'-deoxythymine, 2',3'-dideoxynucleosides, for example, 2'-3'-dideoxyadenosine, 2',3'-dideoxyuridine, 2',3'-dideoxycytosine, 2',3'-dideoxyguanosine, 3'-dideoxyadenosine, 2',3'-dideoxy Examples include, but are not limited to, siuridine, 2',3'-dideoxycytosine, 2',3'-dideoxyguanosine, 2',3'-dideoxythymine, 2'-deoxynucleosides, or chain termination nucleosides such as O-methylnucleosides, 3'-deoxynucleosides, 2',3'-dideoxynucleosides, 3'-O-methylnucleosides, 3'-O-ethylnucleosides, 3'-arabinosides, and other alternative nucleosides known in the art and / or described herein.

[0253] 5.4.5 Secondary structure While not bound by theory, it is intended that stem-loop structures can direct RNA folding, protect the structural stability of nucleic acid molecules (e.g., mRNA), provide recognition sites for RNA-binding proteins, and function as substrates for enzymatic reactions. For example, the incorporation of miR and / or TEE sequences alters the shape of the stem-loop region, increasing and / or decreasing translation (Kedde et al. A Pumilio-induced RNA structure switch in p27-3'UTR controls miR-221 and miR-222 accessibility. Nat Cell Biol., 2010 Oct;12(10):1014-20 (the entire article is incorporated herein by reference)).

[0254] Accordingly, in some embodiments, nucleic acid molecules (e.g., mRNA) or parts thereof described herein may exhibit a stem-loop structure (e.g., histone stem-loops, but not limited to these). In some embodiments, the stem-loop structure is formed from a stem-loop sequence that is about 25 or about 26 nucleotides long (e.g., a sequence described in, but not limited to, International Patent Publication WO2013 / 103659, the entirety of which is incorporated herein by reference). Additional examples of stem-loop sequences are those described in International Patent Publication WO2012 / 019780 and International Patent Publication WO2015 / 02667, the contents of which are incorporated herein by reference. In some embodiments, the stem-loop sequence includes a TEE described herein. In some embodiments, the stem-loop sequence includes a miR sequence described herein. In specific embodiments, the stem-loop sequence may include a miR-122 seed sequence. In a specific embodiment, the nucleic acid molecule includes the stem-loop sequence CAAAGGCTCTTTTCAGAGCCACCA (SEQ ID NO: 47). In another embodiment, the nucleic acid molecule includes the stem-loop sequence CAAAGGCUCUUUUCAGAGCCACCA (SEQ ID NO: 48).

[0255] In some embodiments, the nucleic acid molecule of the Disclosure (e.g., mRNA) includes a stem-loop sequence located upstream (to the 5' end) of the coding region within the nucleic acid molecule. In some embodiments, the stem-loop sequence is located within the 5'-UTR of the nucleic acid molecule. In some embodiments, the nucleic acid molecule of the Disclosure (e.g., mRNA) includes a stem-loop sequence located downstream (to the 3' end) of the coding region within the nucleic acid molecule. In some embodiments, the stem-loop sequence is located within the 3'-UTR of the nucleic acid molecule. In some cases, the nucleic acid molecule may contain two or more stem-loop sequences. In some embodiments, the nucleic acid molecule includes at least one stem-loop sequence in the 5'-UTR and at least one stem-loop sequence in the 3'-UTR.

[0256] In some embodiments, the nucleic acid molecule comprising a stem-loop structure further comprises a stabilizing region. In some embodiments, the stabilizing region comprises at least one chain-ending nucleoside that functions to slow degradation and thus increase the half-life of the nucleic acid molecule. Exemplary chain-ending nucleosides that can be used in connection with this disclosure include 3'-deoxyadenosine (cordycepin), 3'-deoxyuridine, 3'-deoxycytosine, 3'-deoxyguanosine, 3'-deoxythymine, 2',3'-dideoxynucleosides, e.g., 2'-3'-dideoxyadenosine, 2',3'-dideoxyuridine, 2',3'-dideoxycytosine, 2',3'-dideoxyguanosine, 3'-dideoxyadenosine, 2',3'-dideoxy Examples include, but are not limited to, deoxyuridine, 2',3'-dideoxycytosine, 2',3'-dideoxyguanosine, 2',3'-dideoxythymine, 2'-deoxynucleoside, or O-methylnucleoside, 3'-deoxynucleoside, 2',3'-dideoxynucleoside, 3'-O-methylnucleoside, 3'-O-ethylnucleoside, 3'-arabinoside, and other alternative nucleosides known in the art and / or described herein. In other embodiments, the stem-loop structure can be stabilized by modification of the 3'-region of the polynucleotide, which can prevent and / or inhibit the addition of oligio(U) (International Patent Publication WO2013 / 103659, which is incorporated herein by reference in its entirety).

[0257] In some embodiments, the nucleic acid molecules of this disclosure include at least one stem-loop sequence and a poly-A region or polyadenylation signal. Non-limiting examples of polynucleotide sequences including at least one stem-loop sequence and a poly-A region or polyadenylation signal include the polynucleotide sequences described in International Patent Publication WO 2013 / 120497, International Patent Publication WO 2013 / 120629, International Patent Publication WO 2013 / 120500, International Patent Publication WO 2013 / 120627, International Patent Publication WO 2013 / 120498, International Patent Publication WO 2013 / 120626, International Patent Publication WO 2013 / 120499, and International Patent Publication WO 2013 / 120628, the contents of each of these, are incorporated herein by reference in their entirety.

[0258] In some embodiments, nucleic acid molecules comprising a stem-loop sequence and a poly-A region or polyadenylation signal can encode a pathogen antigen or a fragment thereof, such as the polynucleotide sequences described in International Patent Publication WO2013 / 120499 and International Patent Publication WO2013 / 120628, the contents of which are incorporated herein by reference in whole.

[0259] In some embodiments, nucleic acid molecules comprising stem-loop sequences and poly-A regions or polyadenylation signals can encode therapeutic proteins, such as the polynucleotide sequences described in International Patent Publication WO2013 / 120497 and International Patent Publication WO2013 / 120629, the contents of which are incorporated herein by reference in their entirety.

[0260] In some embodiments, nucleic acid molecules comprising a stem-loop sequence and a poly-A region or polyadenylation signal can encode tumor antigens or fragments thereof, such as polynucleotide sequences described in International Patent Publication WO2013 / 120500 and International Patent Publication WO2013 / 120627, the contents of which are incorporated herein by reference in whole.

[0261] In some embodiments, nucleic acid molecules comprising a stem-loop sequence and a poly-A region or polyadenylation signal can encode allergenic or autoimmune antigens, such as polynucleotide sequences described in International Patent Publication WO2013 / 120498 and International Patent Publication WO2013 / 120626, the contents of which are incorporated herein by reference in their entirety.

[0262] 5.4.6 Functional nucleotide analogs In some embodiments, the payload nucleic acid molecules described herein contain only standard nucleotides selected from A (adenosine), G (guanosine), C (cytosine), U (uridine), and T (thymidine). While not theoretically bound, certain functional nucleotide analogs are thought to be able to confer useful properties to nucleic acid molecules. Examples of useful properties in the context of this disclosure include, but are not limited to, increased stability of nucleic acid molecules, reduced immunogenicity of nucleic acid molecules in the induction of innate immune responses, enhanced production of proteins encoded by nucleic acid molecules, increased intracellular delivery and / or retention of nucleic acid molecules, and / or reduced cytotoxicity of nucleic acid molecules.

[0263] Therefore, in some embodiments, the payload nucleic acid molecule comprises at least o...

Claims

1. A protein comprising a variant of the mature glycoprotein E (gE) of varicella-zoster virus (VZV), wherein the variant is (a) (i) cleavage of 37 amino acid residues from the C-terminus of the mature gE, and (ii) amino acid residue substitutions Y569A and Y582G (amino acid residue positions 569 and 582 are the amino acid residue position numbers of the full-length VSV gE); (b) Amino acid residue substitutions Y569A and Y582G (amino acid residue positions 569 and 582 are the amino acid residue position numbers of the full-length VSV gE); (c) Amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A (amino acid residue positions 569, 582, 593, 595, 596, and 598 are the amino acid residue position numbers of the full-length VSV gE); (d) Amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A (amino acid residue positions 582, 593, 595, 596, and 598 are the amino acid residue position numbers of the full-length VSV gE); or (e) (i) the mature gE protein having been cleaved of 50 amino acid residues from the C-terminus, and (ii) the protein comprising the amino acid residue substitution Y569A (where amino acid residue position 569 is the amino acid residue position number of the full-length VSV gE).

2. The protein according to claim 1, wherein the variant comprises the amino acid sequence of SEQ ID NOs: 6, 8, 10, 12, or 3.

3. The protein according to claim 1, wherein the amino acid sequence of the mutant consists of the amino acid sequence of SEQ ID NOs: 6, 8, 10, 12, or 3.

4. The protein according to claim 1, wherein the mutant comprises the amino acid sequence of SEQ ID NO:

6.

5. The protein according to claim 1, wherein the amino acid sequence of the mutant consists of the amino acid sequence of SEQ ID NO:

6.

6. The protein according to claim 1, wherein the variant comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95% identical to SEQ ID NO: 6, 8, 10, or 12.

7. The protein according to claim 1, wherein the variant comprises an amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 6, 8, 10, or 12.

8. The protein according to claim 1, wherein the variant comprises an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95% identical to SEQ ID NO:

6.

9. The protein according to claim 1, wherein the mutant comprises an amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:

6.

10. The protein according to any one of claims 1 to 9, further comprising a VZV gE signal peptide.

11. The protein according to claim 10, wherein the VZV gE signal peptide comprises the amino acid sequence described in SEQ ID NO:

18.

12. The protein according to claim 11, wherein the VZV gE signal peptide comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO:

18.

13. The protein according to any one of claims 1 to 9, further comprising a heterologous signal peptide, wherein the N-terminus of the mutant is fused to the C-terminus of the heterologous signal peptide.

14. The protein according to claim 13, wherein the heterologous signal peptide is a human IgE signal peptide.

15. The protein according to claim 14, wherein the human IgE signal peptide comprises (i) the amino acid sequence described in SEQ ID NO: 23, or (ii) an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO:

23.

16. The protein according to claim 14, wherein the amino acid sequence of the human IgE signal peptide is the amino acid sequence described in SEQ ID NO:

23.

17. The protein according to claim 14, wherein the amino acid sequence of the mutant consists of the amino acid sequence described in SEQ ID NO: 6, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence described in SEQ ID NO:

23.

18. The protein according to claim 14, comprising the amino acid sequence described in Sequence ID No.

59.

19. The protein according to claim 14, wherein the amino acid sequence of the protein consists of the amino acid sequence described in Sequence ID No.

59.

20. The protein according to claim 13, wherein the heterologous signal peptide is a human tPA signal peptide.

21. The protein according to claim 20, wherein the human tPA signal peptide comprises (i) the amino acid sequence described in SEQ ID NO: 27, or (ii) an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence described in SEQ ID NO:

27.

22. The protein according to any one of claims 1 to 21, wherein the mature gE comprises the amino acid sequence described in SEQ ID NO: 1, and / or the full-length VZV gE comprises the amino acid sequence described in SEQ ID NO:

55.

23. A fragment of mature glycoprotein E (gE) of varicella-zoster virus (VZV), wherein the fragment comprises a cleavage of at least one amino acid residue, up to 50, 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus of the mature gE, and optionally, the cleavage comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 The fragment is a cleavage of 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 or an amino acid residue, or optionally, the cleavage is a cleavage of 11, 12, 13, 14, 15, 16, 17, 18, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43 or 44 amino acid residues from the C-terminus of the mature gE, or optionally, the cleavage is a cleavage of 14 or 37 amino acid residues from the C-terminus of the mature gE.

24. The fragment according to claim 23, wherein the fragment further comprises an amino acid residue substitution Y569A, and the amino acid residue position 569 is an amino acid residue position number according to the full-length VZV gE.

25. The fragment according to claim 23 or 24, wherein the fragment comprises cleavage of up to 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, or 31 amino acid residues from the C-terminus of the mature gE.

26. The fragment according to claim 25, wherein the fragment further comprises an amino acid residue substitution Y582G, and amino acid residue position 582 is an amino acid residue position number corresponding to the full-length VZV gE protein.

27. The fragment according to any one of claims 23 to 26, wherein the fragment comprises a cleavage of up to 30 or 29 amino acid residues from the C-terminus of the mature gE.

28. The fragment according to claim 27, wherein the fragment further comprises an amino acid residue substitution S593A, and the amino acid residue position 593 is an amino acid residue position number according to the full-length VZV gE.

29. The fragment according to any one of claims 23 to 28, wherein the fragment comprises cleavage of up to 28 amino acid residues from the C-terminus of the mature gE.

30. The fragment according to claim 29, wherein the fragment includes an amino acid residue substitution S595A, and the amino acid residue position 595 is an amino acid residue position number according to the full-length VSV gE.

31. The fragment according to any one of claims 23 to 30, wherein the fragment comprises a cleavage of up to 27 or 26 amino acid residues from the C-terminus of the mature gE.

32. The fragment according to claim 31, wherein the fragment further comprises an amino acid residue substitution T596A, and the amino acid residue position 596 is an amino acid residue position number according to the full-length VZV gE.

33. The fragment according to any one of claims 23 to 32, wherein the fragment comprises cleavage of up to 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2 amino acid residues, or up to 23 amino acid residues, from the C-terminus of the mature gE.

34. The fragment according to claim 33, wherein the fragment further comprises an amino acid residue substitution T598A, and the amino acid residue position 598 is an amino acid residue position number according to the full-length VZV gE.

35. The fragment according to claim 23, wherein the fragment comprises a cleavage of 37 amino acid residues from the C-terminus of the mature gE.

36. The fragment according to claim 23, wherein the fragment comprises (1) a cleavage of 37 amino acid residues from the C-terminus of the mature gE, and (2) amino acid residue substitutions Y569A and Y582G, wherein amino acid residue positions 569 and 582 are amino acid residue position numbers according to the full-length VZV gE.

37. The fragment according to any one of claims 23 to 36, wherein the mature γE comprises the amino acid sequence described in SEQ ID NO: 1, and / or the full-length VZV γE comprises the amino acid sequence described in SEQ ID NO:

55.

38. The fragment according to any one of claims 23 to 37, wherein the fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence described in SEQ ID NO: 6, 3, 8, 10, or 12.

39. A fusion protein comprising a fragment according to any one of claims 23 to 38 and a heterologous signal peptide, wherein the N-terminus of the fragment is fused to the C-terminus of the heterologous signal peptide.

40. The fusion protein according to claim 39, wherein the heterologous signal peptide is a human tPA signal peptide.

41. The fragment according to claim 40, wherein the human tPA signal peptide comprises the amino acid sequence described in SEQ ID NO:

27.

42. The fusion protein according to claim 39, wherein the heterologous signal peptide is a human IgE signal peptide.

43. The fusion protein according to claim 42, wherein the human IgE signaling peptide comprises the amino acid sequence described in SEQ ID NO:

23.

44. A fusion protein comprising a fragment of mature glycoprotein (gE) of varicella-zoster virus (VZV) and a human IgE signal peptide, wherein the fragment comprises a cleavage of 37 amino acid residues from the C-terminus of the mature gE, and the N-terminus of the fragment is fused to the C-terminus of the human IgE signal peptide.

45. A nucleic acid encoding a protein according to any one of claims 1 to 22, a fragment according to any one of claims 23 to 38, or a fusion protein according to any one of claims 39 to 44.

46. The nucleic acid according to claim 45, comprising a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the nucleotide sequence described in SEQ ID NO: 7, 9, 11, 13, 4, or 5.

47. A nucleic acid encoding a protein according to any one of claims 14 to 19, or a fusion protein according to any one of claims 42 to 44, wherein the nucleotide sequence encoding the IgE signal peptide comprises a nucleotide sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to (a) the nucleotide sequence according to SEQ ID NO: 24, 25, or 26, or (b) the nucleotide sequence according to SEQ ID NO: 24, 25, or 26.

48. A nucleic acid encoding a protein according to any one of claims 10 to 12, wherein the nucleotide sequence of the signal peptide includes (a) the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22, or (b) the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22, which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22.

49. A nucleic acid encoding the protein according to claim 20 or 21, or the fusion protein according to claim 40 or 41, wherein the nucleotide sequence encoding the human tPA signal peptide comprises a nucleotide sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to (a) the nucleotide sequence according to SEQ ID NO: 28, or (b) the nucleotide sequence according to SEQ ID NO:

28.

50. A nucleic acid that does not exist in nature, comprising a coding nucleotide sequence encoding a protein according to any one of claims 1 to 22, a fragment according to any one of claims 23 to 38, or a fusion protein according to any one of claims 39 to 44.

51. The nucleic acid not found in nature according to claim 50, wherein the coding nucleotide sequence is codon-optimized for expression within the target cell, and optionally the target is a non-human mammal or human.

52. (a) a nucleotide sequence described in SEQ ID NO: 7, 9, 11, 13, 4, or 5, or (b) a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 7, 9, 11, 13, 4, or 5, a nucleic acid that does not exist in nature as described in claim 50 or 51.

53. A non-naturally occurring nucleic acid encoding a protein according to any one of claims 14 to 19, or a fusion protein according to any one of claims 42 to 44, wherein the nucleotide sequence encoding the IgE signal peptide comprises (a) the nucleotide sequence according to SEQ ID NO: 24, 25, or 26, or (b) the nucleotide sequence according to SEQ ID NO: 24, 25, or 26, which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence according to SEQ ID NO: 24, 25, or 26.

54. A non-naturally occurring nucleic acid encoding the protein according to any one of claims 10 to 12, wherein the nucleotide sequence of the signal peptide of the full-length VSV gE protein comprises (a) the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22, or (b) the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22, which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 19, 20, 21, or 22.

55. A non-naturally occurring nucleic acid encoding the protein according to claim 20 or 21, or the fusion protein according to claim 40 or 41, wherein the nucleotide sequence encoding the human tPA signal peptide comprises a nucleotide sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to (a) the nucleotide sequence according to SEQ ID NO: 28, or (b) the nucleotide sequence according to SEQ ID NO:

28.

56. Nucleic acids that do not exist in nature and contain the nucleotide sequence of sequence numbers 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5.

57. The nucleic acid according to claim 56, which is not naturally occurring, comprising the nucleotide sequence of sequence numbers 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5.

58. The nucleic acid described above is an unnatural nucleic acid according to any one of claims 50 to 55, which is essentially composed of or includes (1) a nucleotide sequence described in SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5, or (2) a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the nucleotide sequence described in SEQ ID NOs: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5.

59. A nucleic acid that does not exist in nature according to any one of claims 50 to 58, further comprising a 5' untranslated region (5'-UTR) wherein the 5'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 29 to 38, and / or further comprising a 3' untranslated region (3'-UTR) wherein the 3'-UTR comprises a nucleotide sequence described in any one of SEQ ID NOs. 39 to 46, and optionally further comprising a poly A tail or a polyadenylation signal.

60. The nucleic acid that does not exist in nature according to any one of claims 50 to 59, wherein the nucleic acid is DNA.

61. The nucleic acid according to any one of claims 50 to 60, wherein the nucleic acid comprises one or more functional nucleotide analogs.

62. The nucleic acid according to any one of claims 50 to 59, wherein the nucleic acid is mRNA, and thymine is substituted within the nucleic acid with uracil or a functional analog.

63. The nucleic acid according to claim 62, wherein the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methylpsoiduridine, and 5-methylcytosine.

64. The nucleic acid is such that all thymine (T) is replaced with uracil (U) or N1-methylpsoiduridine, and / or the first nucleotide G is m ７ A nucleic acid not found in nature according to claim 62, which comprises, essentially, or includes, a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleotide sequence described in SEQ ID NO: 63, except that it is substituted with GpppAmpU.

65. The nucleic acid is such that all thymine (T) is replaced with uracil (U) or N1-methylpsoiduridine, and / or the first nucleotide G is m ７ A nucleic acid that does not exist in nature according to claim 62, comprising, essentially, or comprising the nucleotide sequence described in SEQ ID NO: 63, except that it is substituted with GpppAmpU.

66. The nucleic acid is such that all thymine (T) is replaced with N1-methylpsoiduridine, and the first nucleotide G is m ７ A nucleic acid that does not exist naturally, according to claim 62, comprising, essentially, or comprising the nucleotide sequence described in SEQ ID NO: 63, except that it is substituted with GpppAmpU.

67. A vector comprising a nucleic acid according to any one of claims 45 to 49 or a nucleic acid not naturally occurring according to any one of claims 50 to 66, preferably an IVT (in vitro transcription) plasmid.

68. A host cell comprising a nucleic acid according to any one of claims 45 to 49, a nucleic acid not naturally occurring according to any one of claims 50 to 66, or a vector according to claim 67.

69. The host cell according to claim 68, which is in vitro, ex vivo, or isolated.

70. A pharmaceutical composition comprising a protein according to any one of claims 1 to 22, a fragment according to any one of claims 23 to 38, or a fusion protein according to any one of claims 39 to 44.

71. A pharmaceutical composition comprising a nucleic acid according to any one of claims 45 to 49, a nucleic acid not found in nature according to any one of claims 50 to 66, or a vector according to claim 67.

72. A pharmaceutical composition comprising a nucleic acid not found in nature as described in any one of claims 50 to 66 and at least a first lipid, wherein the first lipid is optionally a compound according to formula 01-I or formula 01-II, or a compound listed in Table 01-1, or a compound according to formula 02-I, or a compound listed in Table 02-1, or a compound according to formula 03-I, or a compound listed in Table 03-1, or a compound according to formula 04-I, or a compound listed in Table 04-1.

73. The pharmaceutical composition according to claim 72, further comprising a second lipid, wherein the second lipid is optionally a compound according to formula 05-I.

74. The pharmaceutical composition according to claim 72 or 73, formulated as lipid nanoparticles that encapsulate the nucleic acid within a lipid shell.

75. The pharmaceutical composition according to any one of claims 70 to 74, wherein the composition is a vaccine.

76. A method for managing, preventing or treating a disease or disorder in a subject caused by or caused by VZV, comprising administering to the subject a therapeutically effective amount of a protein according to any one of claims 1 to 22, a fragment according to any one of claims 23 to 38, a fusion protein according to any one of claims 39 to 44, a nucleic acid according to any one of claims 45 to 49, a nucleic acid not found in nature according to any one of claims 50 to 66, a vector according to claim 67, or a pharmaceutical composition according to any one of claims 70 to 75.

77. The method according to claim 76, wherein the method is a method for preventing a disease or disorder in a subject caused by or due to VZV.

78. The method according to claim 76 or 77, wherein an immune response to the VZV is induced within the subject.

79. The method according to claim 78, wherein the immune response includes the production of cytokines in lymphocytes.

80. The method according to claim 78 or 79, wherein the immune response includes an increase in the proportion of cytokine-expressing lymphocytes.

81. The aforementioned lymphocytes are CD4 ＋ T cells and / or CD8 ＋ The method according to claim 79 or 80, wherein the T cell and / or the cytokine is one or more of IFN-γ, IL-2, and TNF-α.

82. The method according to any one of claims 79 to 81, wherein the production of the cytokine in lymphocytes is increased.

83. The method according to any one of claims 78 to 82, wherein the immune response includes the production of an antibody that specifically binds to VZV gE.

84. The method according to any one of claims 76 to 83, wherein the disease or disorder caused by VZV is (a) varicella and / or herpes zoster, (b) postherpetic neuralgia (PHN), and / or (c) one or more of meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcer, hepatitis, and pancreatitis.

85. The method according to any one of claims 76 to 84, wherein the subject is a human.

86. The method according to claim 85, wherein the person is an adult human.

87. The method according to claim 86, wherein the adult is at least 40 years old.

88. The method according to claim 85, wherein the person is an elderly person.

89. A protein as defined in any one of claims 1 to 22, a fragment as defined in any one of claims 23 to 38, a fusion protein as defined in any one of claims 39 to 44, a nucleic acid as defined in any one of claims 45 to 49, a nucleic acid not found in nature as defined in any one of claims 50 to 66, a vector as defined in claim 67, or a pharmaceutical composition as defined in any one of claims 70 to 75, for use in a method for managing, preventing or treating a disease or disorder in a subject caused by or due to VZV.

90. The protein, fragment, fusion protein, nucleic acid, nucleic acid not found in nature, vector, or pharmaceutical composition for use according to claim 89, wherein the subject is a human, and optionally an adult or elderly human.

91. Use of a protein as defined in any one of claims 1 to 22, a fragment as defined in any one of claims 23 to 38, a fusion protein as defined in any one of claims 39 to 44, a nucleic acid as defined in any one of claims 45 to 49, a nucleic acid not found in nature as defined in any one of claims 50 to 66, a vector as defined in claim 67, or a pharmaceutical composition as defined in any one of claims 70 to 75, for manufacturing a pharmaceutical for managing, preventing or treating a disease or disorder in a subject caused by or due to VZV.

92. The use according to claim 91, wherein the subject is a human being, and optionally an adult or elderly human being.

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