Mutant fragments of VZV glycoprotein E

Abstract

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

Description

1. Technical Field

[0001] The present disclosure generally relates to fragments of the glycoprotein E (gE) protein of varicella-zoster virus (VZV) and nucleic acid molecules that can be used to control, prevent and treat diseases or conditions caused by VZV or infection with VZV. The disclosure also relates to lipid-containing compositions (including vaccines) containing the nucleic acid molecules. 2. Background Technology

[0002] Varicella zoster virus (VZV), also known as human herpesvirus type 3, is a double-stranded DNA virus belonging to alpha herpesvirus. VZV has only one serotype. VZV has a genome comprising 71 genes and encoding 67 different proteins, including 6 glycoproteins, which are now 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 envelope of virions. Antibodies induced by the three major glycoproteins can neutralize the virus. Specific humoral and cellular immunity and cytokines such as interferon play a major role in limiting the spread and recovery of VZV, wherein specific cellular immunity is particularly important.

[0003] Zostavax (MSD) is a live attenuated virus vaccine. It reduces viral load by 61.1% (65.5% in people aged 60-69 years and 55.4% in people aged 70 years and older). It also reduces the duration of pain and discomfort caused by the virus. Shinrix (GSK) is a subunit vaccine. It contains gE and AS01B, an adjuvant system used to enhance cellular immune responses. It can achieve efficacy exceeding 90%. 3. Summary of the Invention

[0004] In one aspect, provided herein are fragments of the gE protein of VZV, optionally comprising a mutation, such as a substitution.

[0005] In some embodiments, the fragment comprises a truncation of at least 1 amino acid residue and up to 49 amino acid residues from the C-terminus compared to the mature gE protein. In some embodiments, the fragment comprises a truncation 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 1 amino acid residue from the C-terminus compared to the mature gE protein. In some embodiments, the fragment comprises a truncation 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 some embodiments, the fragment comprises a truncation 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 the mature gE protein. In some embodiments, the fragment comprises a truncation of 14 or 37 amino acid residues from the C-terminus compared to the mature gE protein.

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

[0007] In some embodiments, the fragment comprises a truncation 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 comprises the substitution Y569A.

[0008] In some embodiments, the fragment comprises a truncation 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 fragment optionally comprises the substitution Y582G, with or without the substitution Y569A. In such embodiments, the fragment comprises the substitutions Y569A and Y582G.

[0009] In some embodiments, compared with mature gE protein, the fragment comprises a truncation of 30 or 29 amino acid residues from the C-terminus. In such embodiments, the fragment optionally comprises a substitution of S593A, with or without substitutions of Y569A and / or Y582G. In such embodiments, the fragment comprises a substitution of Y569A, Y582G and S593A. In some embodiments, compared with mature gE protein, the fragment comprises a truncation of 28 amino acid residues from the C-terminus. In such embodiments, the fragment optionally comprises a substitution of S595A, with or without substitutions of Y569A and / or Y582G and / or S593A. In such embodiments, the fragment comprises a substitution of Y569A, Y582G, S593A and S595A. In some embodiments, compared with mature gE protein, the fragment comprises a truncation of 27 or 26 amino acid residues from the C-terminus. In such embodiments, the fragment optionally comprises replacement T596A, has or does not replace Y569A and / or Y582G and / or S593A and / or S595A. In such embodiments, the fragment comprises replacement Y569A, Y582G, S593A, S595A and T596A. In some embodiments, compared with mature gE protein, the fragment comprises from C-terminal 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 the truncation of 1 amino acid residue. In such embodiments, the fragment optionally comprises replacement T598A, has or does not replace Y569A and / or Y582G and / or S593A and / or S595A and / or T596A. In such embodiments, the fragment comprises the substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A.

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

[0011] In some embodiments, the fragment comprises the amino acid sequence shown in SEQ ID NO:3, 6, 8, 10 or 12, or an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO:3, 6, 8, 10 or 12.

[0012] In some embodiments, the N-terminus of the fragment is fused to the C-terminus of a 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 comprises the amino acid sequence set forth 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 comprises the amino acid sequence set forth 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 comprises the amino acid sequence set forth in SEQ ID NO: 23.

[0013] In one aspect, provided herein are nucleic acids encoding the fragments as described herein.

[0014] In some embodiments, the fragment is encoded by the nucleotide sequence shown in SEQ ID NO:4, 5, 7, 9, 11 or 13, or 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 shown in SEQ ID NO:4, 5, 7, 9, 11 or 13.

[0015] In some embodiments, the native signal peptide of the gE protein of VZV is encoded by the nucleotide sequence set forth in SEQ ID NO: 19, 20, 21, or 22, or 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 set forth in SEQ ID NO: 19, 20, 21, or 22. In some embodiments, the human tPA signal peptide is encoded by the nucleotide sequence set forth in SEQ ID NO: 28, or 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 set forth in SEQ ID NO: 28. In some embodiments, the human IgE signal peptide is encoded by the nucleotide sequence shown in SEQ ID NO: 24, 25 or 26, or 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 shown in SEQ ID NO: 24, 25 or 26.

[0016] In some embodiments, provided herein is a protein comprising a mutant of mature glycoprotein E (gE) of varicella-zoster virus (VZV), wherein the mutant comprises: (a) (i) a truncation of 37 amino acid residues from the C-terminus of mature gE, and (ii) amino acid residue substitutions Y569A and Y582G, wherein amino acid residue positions 569 and 582 are the amino acid residue position numbering of full-length VSV gE; (b) amino acid residue substitutions Y569A and Y582G, wherein amino acid residue positions 569 and 582 are the amino acid residue position numbering of full-length VSV gE. (c) amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residue positions 569, 582, 593, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VSV gE; (d) amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residue positions 582, 593, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VSV gE; or (e) (i) a truncation of the C-terminal 50 amino acid residues of the mature gE protein, and (ii) an amino acid residue substitution Y569A, wherein amino acid residue position 569 is numbered according to the amino acid residue position of full-length VSV gE. In some embodiments, the mutant comprises the amino acid sequence of SEQ ID NO: 6, 8, 10, 12, or 3. In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence of SEQ ID NO: 6, 8, 10, 12, or 3. In some embodiments, the mutant comprises the amino acid sequence of SEQ ID NO: 6. In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence of SEQ ID NO: 6. In some embodiments, the mutant 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. In some embodiments, 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, 8, 10, or 12. In some embodiments, the mutant 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. In some embodiments, 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. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55.In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO:1, and the full-length VZV gE comprises the amino acid sequence set forth 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 set forth 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 set forth in SEQ ID NO:18. In some embodiments, the protein further comprises a heterologous signal peptide, wherein the N-terminus of the mutant 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 the amino acid sequence set forth 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 set forth in SEQ ID NO:27. In some embodiments, the amino acid sequence of the human tPA signal peptide consists of the amino acid sequence set forth 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 set forth 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 set forth in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO: 23. In some embodiments, the protein comprises the amino acid sequence set forth in SEQ ID NO: 59.

[0018] In some embodiments, provided herein is a protein comprising a mutant of mature gE of VZV and a human IgE signal peptide, wherein the mutant comprises the amino acid sequence shown in SEQ ID NO: 6, and wherein the amino acid sequence of the human IgE signal peptide comprises the amino acid sequence shown in SEQ ID NO: 23. In some embodiments, provided herein is a protein comprising a mutant of mature gE of VZV and a human IgE signal peptide, wherein the amino acid sequence of the mutant consists of the amino acid sequence shown in SEQ ID NO: 6, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence shown in SEQ ID NO: 23. In some embodiments, provided herein is a protein consisting of a mutant of mature gE of VZV and a human IgE signal peptide, wherein the amino acid sequence of the mutant consists of the amino acid sequence shown in SEQ ID NO: 6, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence shown in SEQ ID NO: 23.

[0019] In some embodiments, provided herein is a protein comprising the amino acid sequence shown in SEQ ID NO: 59. In some embodiments, 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 shown in SEQ ID NO: 59. In some embodiments, 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 shown in SEQ ID NO: 59. In some embodiments, provided herein is a protein comprising 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 shown in SEQ ID NO: 59. In some embodiments, provided herein is a protein, wherein the amino acid sequence of the protein consists of the amino acid sequence shown in SEQ ID NO: 59.

[0020] In some embodiments, provided herein are fragments of mature gE of VZV, wherein the fragment comprises a truncation of at least one and at most 50, 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus of mature gE. In some embodiments, the truncation is a truncation 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 truncation is a truncation 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 truncation is a truncation 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, wherein the amino acid position numbering is based on the full-length VZV gE. In some embodiments, the fragment further comprises the amino acid residue substitution Y569A, and wherein amino acid residue position 569 is numbered according to the amino acid residue position of the full-length VZV gE. In some embodiments, the fragment comprises 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 shown in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment comprises 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 shown in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment 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 shown in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment 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 set forth in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment comprises the amino acid sequence set forth 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 shown in SEQ ID NO: 6, 3, 8, 10, or 12. In some embodiments, the fragment comprises the amino acid sequence shown in SEQ ID NO: 6. In some embodiments, the amino acid sequence of the fragment consists of the amino acid sequence shown in SEQ ID NO: 6.

[0021] In some embodiments, provided herein are fragments of mature gE of VZV, wherein the fragment comprises a truncation 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, wherein amino acid position numbering is based on full-length VZV gE. In some embodiments, the fragment further comprises the amino acid residue substitution Y582G, and wherein amino acid residue position 582 is numbered based on amino acid residue position numbering of the full-length VZV gE protein. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO:1, and the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO:55.

[0022] In some embodiments, provided herein are fragments of mature gE of VZV, wherein the fragment comprises a truncation 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, wherein the amino acid position numbering is based on the full-length VZV gE. In some embodiments, the fragment further comprises the amino acid residue substitution S593A, and wherein amino acid residue position 593 is numbered based on the amino acid residue position numbering of the full-length VZV gE. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1, and full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55.

[0023] In some embodiments, provided herein are fragments of mature gE of VZV, wherein the fragment comprises a truncation 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, wherein amino acid position numbering is based on full-length VZV gE. In some embodiments, the fragment comprises the amino acid residue substitution S595A, and wherein amino acid residue position 595 is numbered based on amino acid residue position numbering of full-length VZV gE. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1, and full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55.

[0024] In some embodiments, provided herein are fragments of mature gE of VZV, wherein the fragment comprises a truncation 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, wherein the amino acid position numbering is based on the full-length VZV gE. In some embodiments, the fragment further comprises the amino acid residue substitution T596A, and wherein amino acid residue position 596 is numbered based on the amino acid residue position numbering of the full-length VZV gE. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1, and full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55.

[0025] In some embodiments, provided herein are fragments of mature gE of VZV, wherein the fragment comprises 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 truncation 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, wherein the amino acid position numbering is based on the full-length VZV gE. In some embodiments, the fragment further comprises the amino acid residue substitution T598A, and wherein amino acid residue position 598 is numbered according to the amino acid residue position of the full-length VZV gE. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1, and full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55.

[0026] In some embodiments, provided herein are fragments of mature gE of VZV, wherein the fragment comprises a truncation of the C-terminal 37 amino acid residues 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, wherein the amino acid position numbering is based on the full-length VZV gE. In some embodiments, the fragment comprises: (1) a truncation of the C-terminal 37 amino acid residues of mature gE, and (2) amino acid residue substitutions Y569A and Y582G, wherein amino acid residue positions 569 and 582 are numbered according to the amino acid residue positions of the full-length VZV gE. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO:1, and the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO:55.

[0027] 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 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 set forth in SEQ ID NO:28. In some embodiments, the human tPA signal peptide comprises the amino acid sequence set forth 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 set forth 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 set forth in SEQ ID NO:23. In some embodiments, the fragment comprises a truncation of at least 1 and at most 50, 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus of mature gE. In some embodiments, the truncation is a truncation 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 truncation is a truncation 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 truncation is a truncation 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, wherein the amino acid position numbering is based on the full-length VZV gE. In some embodiments, the fragment is a fragment described herein. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO:1, and the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO:55.

[0028] In some embodiments, the present invention provides a fusion protein comprising a fragment of mature gE of VZV and a human IgE signal peptide, wherein the fragment comprises a truncation 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 shown in SEQ ID NO: 23. In some embodiments, the human IgE signal peptide comprises the amino acid sequence shown in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence shown in SEQ ID NO: 23. In some embodiments, mature gE comprises the amino acid sequence shown in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence shown in SEQ ID NO: 55. In some embodiments, mature gE comprises the amino acid sequence shown in SEQ ID NO: 1, and full-length VZV gE comprises the amino acid sequence shown in SEQ ID NO: 55.

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

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

[0031] In some embodiments, provided herein is a nucleic acid encoding a protein described herein, wherein the protein comprises a mutant of mature gE of VZV and a human IgE signal peptide, and wherein the nucleotide sequence encoding the IgE signal peptide comprises the nucleotide sequence shown in SEQ ID NO: 24, 25, or 26. In some embodiments, provided herein is a nucleic acid encoding a protein described herein, wherein the protein comprises a mutant of mature gE of VZV and a human IgE signal peptide, and wherein the nucleotide sequence encoding the IgE 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 shown 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, wherein the fusion protein comprises a fragment of mature gE of VZV and a human IgE signal peptide, and wherein the nucleotide sequence encoding the IgE signal peptide comprises the nucleotide sequence shown in SEQ ID NO: 24, 25, or 26. In some embodiments, provided herein is a nucleic acid encoding a fusion protein as described herein, wherein the fusion protein comprises a fragment of mature gE of VZV and a human IgE signal peptide, and wherein the nucleotide sequence encoding the IgE 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 shown in SEQ ID NO: 24, 25, or 26.

[0033] In some embodiments, provided herein is a nucleic acid encoding a protein described herein, wherein the protein comprises a mutant of mature gE of VZV and a VZV gE signal peptide, wherein the nucleotide sequence of the signal peptide comprises the nucleotide sequence shown in SEQ ID NO: 19, 20, 21, or 22. In some embodiments, provided herein is a nucleic acid encoding a protein described herein, wherein the protein comprises a mutant of mature gE of VZV and a VZV gE signal peptide, wherein the nucleotide sequence of the 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 shown in SEQ ID NO: 19, 20, 21, or 22.

[0034] In some embodiments, provided herein is a nucleic acid encoding a protein described herein, wherein the protein comprises a mutant 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 shown in SEQ ID NO: 28. In some embodiments, provided herein is a nucleic acid encoding a protein described herein, wherein the protein comprises 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 shown in SEQ ID NO: 28.

[0035] In some embodiments, provided herein is a nucleic acid encoding a fusion protein as described herein, wherein the fusion protein comprises 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 shown in SEQ ID NO: 28. In some embodiments, provided herein is a nucleic acid encoding a fusion protein as described herein, wherein the fusion protein comprises 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 shown in SEQ ID NO: 28.

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

[0037] In one aspect, provided herein are non-naturally occurring nucleic acid molecules that can be used to prevent, manage, and treat diseases or conditions caused by VZV or infection with VZV.

[0038] In some embodiments, the non-naturally occurring nucleic acid comprises a nucleotide sequence encoding a fragment as described herein. In some embodiments, the fragment consists of, consists essentially of, or comprises 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 shown in SEQ ID NO: 1 or a fragment thereof. In some embodiments, the fragment consists of, consists essentially 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 to the amino acid sequence shown in SEQ ID NO: 3, 6, 8, 10, or 12. In some embodiments, the fragment consists of, consists essentially of, or comprises an amino acid sequence shown in SEQ ID NO: 3, 6, 8, 10, or 12. In some embodiments, the encoding nucleotide sequence consists of, consists essentially of, or comprises a nucleotide sequence that has at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleotide sequence shown in SEQ ID NO: 2, or a fragment thereof. In some embodiments, the encoding nucleotide sequence has been codon-optimized for expression in a subject's cells. In some embodiments, the subject is a non-human mammal. In some embodiments, the subject is a human. In some embodiments, the encoding nucleotide sequence consists of, consists essentially of, or comprises 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 to the nucleotide sequence shown in SEQ ID NO: 4, 5, 7, 9, 11, or 13. In some embodiments, the encoding nucleotide sequence consists of, consists essentially of, or comprises the nucleotide sequence shown in SEQ ID NO: 4, 5, 7, 9, 11, or 13. In some embodiments, the fragment is fused to the gE native signal peptide. In some embodiments, the signal peptide consists of, consists essentially 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 to the amino acid sequence shown in SEQ ID NO:18.In some embodiments, the signal peptide is encoded by a nucleotide sequence consisting of, consisting essentially 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 to the nucleotide sequence shown 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 a human immunoglobulin, a signal peptide, and a peptide that promotes multimerization of the fusion protein. In some embodiments, the signal peptide is a signal peptide from IgE or tPA. In some embodiments, the signal peptide consists of, consists essentially 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 to the amino acid sequence shown in SEQ ID NO: 23. In some embodiments, the signal peptide is encoded by a coding nucleotide sequence consisting of, consists essentially of, or comprises 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 to the nucleotide sequence shown in SEQ ID NO: 24, 25, or 26. In some embodiments, the signal peptide consists of, consists essentially 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 to the amino acid sequence shown in SEQ ID NO: 27. In some embodiments, the signal peptide is encoded by a coding nucleotide sequence consisting of, consists essentially of, or comprises 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 to the nucleotide sequence shown in SEQ ID NO: 28. In some embodiments, multimerization is dimerization or trimerization. In some embodiments, the non-naturally occurring nucleic acid further comprises a 5' untranslated region (5'-UTR), wherein the 5'-UTR comprises the sequence shown in any one of SEQ ID NOs: 29-38. In some embodiments, the non-naturally occurring nucleic acid further comprises a 3' untranslated region (3'-UTR), wherein the 3'-UTR comprises the sequence shown 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 consists of, consists essentially of, or comprises the nucleotide sequence shown in SEQ ID NO: 49, 50, 51, 52, 53, 54, 60, 61, 62, 63, or 64, 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 shown in SEQ ID NO: 49, 50, 51, 52, 53, 54, 60, 61, 62, 63, or 64. In some embodiments, the non-naturally occurring nucleic acid comprises one or more functional nucleotide analogs. In some embodiments, the non-naturally occurring nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine (psd), 1-methyl-pseudouridine (m1), and 5-methylcytosine. In some embodiments, the nucleic acid is DNA or mRNA. In some embodiments, the nucleic acid consists of, consists essentially of, or comprises the nucleotide sequence shown 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 shown in SEQ ID NO:63, wherein all thymines (T) are replaced by uracil (U) or N1-methylpseudouridine, and / or the first nucleotide G is replaced by m. 7 GpppAmpU replaced.

[0039] In some embodiments, disclosed herein are vectors or cells comprising a non-naturally occurring nucleic acid molecule as described herein. In some embodiments, the vector is preferably an IVT plasmid. In some embodiments, disclosed herein are compositions comprising a non-naturally occurring nucleic acid molecule as described herein.

[0040] In some embodiments, a non-naturally occurring nucleic acid is provided herein, comprising a coding nucleotide sequence encoding a protein described herein. In some embodiments, a non-naturally occurring nucleic acid is provided herein, comprising a coding nucleotide sequence encoding a fragment described herein. In some embodiments, a non-naturally occurring nucleic acid is provided herein, comprising a coding nucleotide sequence encoding a fusion protein described herein. In some embodiments, the coding nucleotide sequence has been codon-optimized for expression in a subject's cells. In some embodiments, the coding nucleotide sequence has been codon-optimized for expression in non-human mammalian cells. In specific embodiments, the coding nucleotide sequence has been codon-optimized for expression in human cells. In some embodiments, the non-naturally occurring nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 7, 9, 11, 13, 4, or 5. The non-naturally occurring nucleic acid 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 shown in SEQ ID NO: 7, 9, 11, 13, 4, or 5. In some embodiments, the non-naturally occurring nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 7.

[0041] In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding a protein described herein, wherein the protein comprises a mutant of mature gE of VZV and a human IgE signal peptide, and wherein the nucleotide sequence encoding the IgE signal peptide comprises the nucleotide sequence shown in SEQ ID NO: 24, 25, or 26. In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding a protein described herein, wherein the protein comprises a mutant of mature gE of VZV and a human IgE signal peptide, and wherein the nucleotide sequence encoding the IgE 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 shown in SEQ ID NO: 24, 25, or 26.

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

[0043] In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding a protein described herein, wherein the protein comprises a mutant of mature gE of VZV and a VZV gE signal peptide, wherein the nucleotide sequence of the signal peptide comprises the nucleotide sequence shown in SEQ ID NO: 19, 20, 21, or 22. In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding a protein described herein, wherein the protein comprises a mutant of mature gE of VZV and a VZV gE signal peptide, wherein the nucleotide sequence of the 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 shown in SEQ ID NO: 19, 20, 21, or 22.

[0044] In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding a protein described herein, wherein the protein comprises a mutant 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 shown in SEQ ID NO: 28. In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding a protein described herein, wherein the protein comprises 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 shown in SEQ ID NO: 28.

[0045] In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding a fusion protein described herein, wherein the fusion protein comprises 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 shown in SEQ ID NO: 28. In some embodiments, provided herein is a non-naturally occurring nucleic acid encoding a fusion protein described herein, wherein the fusion protein comprises 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 shown in SEQ ID NO: 28.

[0046] In some embodiments, provided herein is a non-naturally occurring nucleic acid comprising the nucleotide sequence of SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, the non-naturally occurring nucleic acid consists of the nucleotide sequence of SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, the non-naturally occurring nucleic acid consists of, consists essentially of, or comprises the nucleotide sequence shown in SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, the non-naturally occurring nucleic acid 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 shown in SEQ ID NO:63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, the non-naturally occurring nucleic acid consists of, consists essentially of, or 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 shown in SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, the non-naturally occurring nucleic acid consists of, consists essentially of, or comprises the nucleotide sequence shown in SEQ ID NO: 63, 51, 60, 62, 62, or 64. In some embodiments, the non-naturally occurring nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 63. In some embodiments, the non-naturally occurring nucleic acid consists of the nucleotide sequence shown in SEQ ID NO: 63.

[0047] In some embodiments, the non-naturally occurring nucleic acids described herein further comprise a 5' untranslated region (5'-UTR) and / or a 3' untranslated region (3'-UTR). In some embodiments, the non-naturally occurring nucleic acids described herein further comprise a 5'-UTR, wherein the 5'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 29-38. In some embodiments, the non-naturally occurring nucleic acids described herein further comprise a 3'-UTR, wherein the 3'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 39-46. In some embodiments, the non-naturally occurring nucleic acids described herein further comprise a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 29-38, and wherein the 3'-UTR comprises the nucleotide sequence shown 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.

[0048] In some embodiments, the non-natural nucleic acid described herein comprises DNA. In some embodiments, the non-natural nucleic acid described herein is DNA. In some embodiments, the non-natural nucleic acid described herein comprises one or more functional nucleotide analogs. In some embodiments, the non-natural nucleic acid described herein comprises mRNA, and wherein thymine is replaced by uracil or its functional analog in nucleic acid. In some embodiments, the non-natural nucleic acid described herein is mRNA, and wherein thymine is replaced by uracil or its functional analog in nucleic acid. In some embodiments, the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine and 5-methylcytosine. In some embodiments, the nucleic acid is composed of, essentially composed of or comprises the nucleotide sequence shown in SEQ ID NO:63, except that all thymines (T) are replaced by uracil (U) or N1-methylpseudouridine. In some embodiments, the nucleic acid consists of, consists essentially of, or comprises the nucleotide sequence shown in SEQ ID NO: 63, except that all thymines (T) are replaced by uracil (U) or N1-methylpseudouridine, and the first nucleotide G is replaced by m 7GpppAmpU is replaced. In some embodiments, the nucleic acid consists of, consists essentially of, or 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 shown in SEQ ID NO: 63; except that all thymines (T) are replaced by uracils (U) or N1-methylpseudouridines. In some embodiments, the nucleic acid consists of, consists essentially of, or 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 shown in SEQ ID NO: 63; and the first nucleotide G is replaced by m 7 In some embodiments, the nucleic acid consists of, consists essentially of, or 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 shown in SEQ ID NO: 63, except that all thymines (T) are replaced by uracils (U) or N1-methylpseudouridines, and the first nucleotide G is replaced by m 7 In some embodiments, the nucleic acid consists of, consists essentially of, or comprises the nucleotide sequence shown in SEQ ID NO: 63, except that all thymines (T) are replaced by N1-methylpseudouridine and the first nucleotide G is replaced by m 7 GpppAmpU replaced.

[0049] In some embodiments, provided herein is a vector comprising a nucleic acid as described herein. In some embodiments, provided herein is a vector comprising a non-naturally occurring nucleic acid as described herein. In some embodiments, the vector is an IVT (in vitro transcription) plasmid.

[0050] In some embodiments, a host cell is provided herein, comprising a nucleic acid as described herein. In some embodiments, a host cell is provided herein, comprising a non-naturally occurring nucleic acid as described herein. In some embodiments, a host cell is provided herein, comprising a vector as described herein. In some embodiments, the host cell expresses a protein as described herein, a fragment as described herein, or a fusion protein as described herein. In some embodiments, the host cell is in vitro, ex vivo, or isolated.

[0051] In some embodiments of compositions as described herein, compositions also include at least one lipid as described herein.In some embodiments of compositions as described herein, compositions also include at least the first lipid as described herein (e.g., a cationic lipid) and an optional second lipid as described herein (e.g., a polymer-bound 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 a lipid nanoparticle that encapsulates the nucleic acid in a lipid shell. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the composition is a vaccine.

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

[0055] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a nucleic acid as described herein is provided herein. In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a non-naturally occurring nucleic acid as described herein is provided herein. In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a vector as described herein is provided herein. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the pharmaceutical composition is a vaccine. In some embodiments, the nucleic acid as described herein, the non-naturally occurring nucleic acid, or the vector as described herein is used to immunize a subject against VZV. In some embodiments, the nucleic acid as described herein, the non-naturally occurring nucleic acid as described herein, or the vector as described herein is used to induce an immune response in a subject (e.g., such as the immune response described in, for example, Section 6). In some embodiments, the immune response comprises a humoral immune response against VZV. In some embodiments, the immune response comprises a cellular response against VZV. In some embodiments, the immune response comprises antibodies (e.g., neutralizing antibodies) specific for VZV gE.

[0056] In some embodiments, provided herein is a pharmaceutical composition comprising a non-naturally occurring nucleic acid 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 of claim 72 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 a lipid nanoparticle encapsulating the nucleic acid in a lipid shell. In some embodiments, the pharmaceutical composition is a vaccine.

[0057] In one aspect, provided herein are methods for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject, 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 non-naturally occurring nucleic acid described herein, or a therapeutically effective amount of a pharmaceutical composition as described herein.

[0058] In some embodiments of the methods described herein, the subject is a human or non-human mammal. In some embodiments, the subject is a human adult, a human child, or a human toddler. In some embodiments, the subject has a disease or condition. In some embodiments, the subject is at risk for or susceptible to VZV infection. In some embodiments, the subject is elderly. In some embodiments, the subject has been diagnosed as positive for VZV infection. In some embodiments, the subject is asymptomatic.

[0059] In some embodiments of the methods described herein, the methods comprise administering to a subject lipid nanoparticles encapsulating a nucleic acid, and wherein the lipid nanoparticles are endocytosed by cells in the subject. In some embodiments, the nucleic acid is expressed by cells in the subject.

[0060] In some embodiments of the methods described herein, an immune response against VZV is elicited in a subject. In some embodiments, the immune response comprises the production of cytokines in lymphocytes. In some embodiments, the immune response comprises an increase in the proportion of lymphocytes that express cytokines. In some embodiments, the lymphocytes are CD4 + T cells and / or CD8 + In some embodiments, the cytokine is one or more of IFN-γ, IL-2, and TNF-α. In some embodiments, cytokine production in lymphocytes is increased. In some embodiments, the immune response includes 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 by VZV. In some embodiments, the serum titer of the antibody is increased in the subject.

[0061] In some embodiments of the methods described herein, the antibody binds to viral particles or infected cells and marks the viral particles of the infected cells for destruction by the subject's immune system. In some embodiments, endocytosis of viral particles bound by the antibody is induced or enhanced. In some embodiments, antibody-dependent cell-mediated cytotoxicity (ADCC) for infected cells in a subject is induced or enhanced. In some embodiments, antibody-dependent cellular phagocytosis (ADCP) for infected cells in a subject is induced or enhanced. In some embodiments, complement-dependent cytotoxicity (CDC) for infected cells in a subject is induced or enhanced.

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

[0063] In some embodiments, provided herein is a method for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject, the method comprising administering to the subject a therapeutically effective amount of a protein as described herein. In some embodiments, provided herein is a method for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject, the method comprising administering to the subject a therapeutically effective amount of a fragment as described herein. In some embodiments, provided herein is a method for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject, the method comprising administering to the subject a therapeutically effective amount of a fusion protein as described herein. In some embodiments, provided herein is a method for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject, the method comprising administering to the subject a therapeutically effective amount of a nucleic acid as described herein. In some embodiments, provided herein is a method for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject, the method comprising administering to the subject a therapeutically effective amount of a non-naturally occurring nucleic acid as described herein. In some embodiments, provided herein is a method for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject, the method comprising administering to the subject a therapeutically effective amount of a vector described herein. In some embodiments, provided herein is a method for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition described herein. In some embodiments, the method is used to prevent a disease or condition caused by VZV or infection with VZV in a subject. In some embodiments, an immune response against VZV is elicited in the subject. In some embodiments, the immune response comprises the production of cytokines in lymphocytes. In some embodiments, the immune response comprises an increase in the proportion of lymphocytes that express cytokines. In some embodiments, the lymphocytes are CD4 + T cells and / or CD8 +T cells, and / or, wherein the cytokine is one or more of IFN-γ, IL-2 and TNF-α. In some embodiments, the production of cytokines in lymphocytes is increased. In some embodiments, the immune response includes the production of antibodies (e.g., neutralizing antibodies) that specifically bind to VZV gE. In some embodiments, the disease or condition 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 condition 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, ulcer, hepatitis and pancreatitis. In some embodiments, the disease or condition caused by VZV is one or more of (a) varicella or herpes zoster; (b) postherpetic neuralgia (PHN); and (c) meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcer, hepatitis, and pancreatitis. In some embodiments, the disease or condition caused by VZV is one or more of (a) varicella and herpes zoster; (b) postherpetic neuralgia (PHN); and (c) meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcer, hepatitis, and pancreatitis. In some embodiments, the subject is a human. In some embodiments, the human is a human adult. 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 person.

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

[0065] In some embodiments, provided herein are uses of proteins described herein, fragments described herein, fusion proteins described herein for the manufacture of a medicament for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject. In some embodiments, provided herein are uses of nucleic acids described herein, non-naturally occurring nucleic acids described herein, or vectors described herein for the manufacture of a medicament for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject. In some embodiments, provided herein are uses of pharmaceutical compositions described herein for the manufacture of a medicament for controlling, preventing, or treating a disease or condition caused by VZV or infection with VZV in a subject. In some embodiments, the disease or condition caused by VZV is one or more of (a) varicella or herpes zoster; (b) postherpetic neuralgia (PHN); or (c) meningoencephalitis, myelitis, cranial nerve palsy, vasculopathy, keratitis, retinopathy, ulcer, hepatitis, and pancreatitis. In some embodiments, the disease or condition 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, ulcer, hepatitis, and pancreatitis. In some embodiments, the disease or condition 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, ulcer, hepatitis, and pancreatitis. In some embodiments, the disease or condition 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, ulcer, hepatitis, and pancreatitis. In some embodiments, the subject is human. In some embodiments, the human is a human adult. In some embodiments, an adult is at least 40 years old. In some embodiments, an adult is at least 45 years old. In some embodiments, an adult is at least 50 years old. In some embodiments, an adult is at least 55 years old. In some embodiments, an adult is at least 60 years old. In some embodiments, the human is an elderly person. 4. Description of the Figures

[0066] Figure 1 In vitro expression of candidates by transfected HEK293T cells is shown. The suffix psd indicates pseudo-U modification, and the suffix ml indicates 1-N-pseudo-U modification.

[0067] Figure 2 The gE-specific IgG titers induced by the candidates are shown. The suffix psd indicates a pseudo-U modification, and the suffix ml indicates a 1-N-pseudo-U modification.

[0068] Figure 3 gE peptide pool-specific T cell responses induced by the candidates are shown. The suffix psd indicates pseudo-U modification, and the suffix ml indicates 1-N-pseudo-U modification.

[0069] Figure 4 The predicted secondary structure of full-length VZV gE is shown. A, wild type; B, mutant containing substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A.

[0070] Figure 5 The median fluorescence intensity of cells transfected in vitro with the candidates is shown.

[0071] Figure 6 The expression rates of the candidates in in vitro transfected cells are shown.

[0072] Figure 7 Shown are gE-specific IgG titers induced by the candidates.

[0073] Figure 8 Shown are gE peptide pool-specific T cell responses induced by the candidates.

[0074] Figure 9 The location of the target protein in the cell is shown. Green represents the gE protein, red represents the Golgi apparatus, and blue represents the cell nucleus.

[0075] Figure 10 Shown is the analysis of the inhibition of inflammatory responses through RIG-I activation and IFN-β release in vitro using mRNA.

[0076] Figure 11 The expression rates of the candidates in in vitro transfected cells are shown.

[0077] Figure 12 The median fluorescence intensity of cells transfected in vitro with the candidates is shown.

[0078] Figure 13A Shown are gE-specific IgG titers after the first immunization in naive mice.

[0079] Figure 13B Shown are gE-specific IgG titers after boosting in naive mice.

[0080] Figure 13C Shown are gE-specific IgG titers after the first immunization in LAV-experienced mice.

[0081] Figure 13D Shown are gE-specific IgG titers following boosting in LAV-experienced mice.

[0082] Figure 14A Shown are CD4+ T cell responses induced by the vaccine in naive mice.

[0083] Figure 14B Shown are vaccine-induced CD4+ T cell responses in LAV-experienced mice. 5. Specific implementation methods

[0084] Provided herein are therapeutic nucleic acid molecules that can be used to prevent, control, and treat diseases or conditions caused by VZV or infection with VZV. Also provided herein are pharmaceutical compositions comprising the therapeutic nucleic acid molecules, including pharmaceutical compositions formulated as lipid nanoparticles, and related treatment methods and uses for preventing, controlling, and treating diseases or conditions caused by VZV or infection with VZV. Additional features of the present disclosure will become apparent to those skilled in the art upon consideration of the following detailed description of specific embodiments.

[0085] 5.1 General Technology

[0086] The techniques and procedures described or referenced herein include those generally well understood and / or commonly employed by those skilled in the art using routine methods, such as the widely used methods described in Sambrook et al., Molecular Cloning: A Laboratory Manual (3rd ed., 2001); Current Protocols in Molecular Biology (Ausubel et al., eds., 2003).

[0087] 5.2 Terminology

[0088] Unless otherwise described, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. For the purpose of interpreting this specification, the following terminology will be applied, and where appropriate, terms used in the singular will also include the plural form, and vice versa. All patents, applications, published applications and other publications are incorporated by reference in their entirety. If any description of a term set forth conflicts with any document incorporated herein by reference, the terminology set forth below will prevail.

[0089] As used herein and unless otherwise indicated, the term "lipid" refers to a group of organic compounds that include, but are not limited to, esters of fatty acids and are generally characterized by being poorly soluble in water but soluble in many non-polar organic solvents. Although lipids are generally poorly soluble in water, certain classes of lipids (e.g., lipids modified with polar groups, such as DMG-PEG2000) have limited water solubility and can be soluble 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 categories: (1) "simple lipids," which include fats and oils as well as waxes; (2) "complex lipids," which include phospholipids and glycolipids (e.g., DMPE-PEG2000); and (3) "derivatized lipids," such as steroids. Further, as used herein, lipids also encompass lipid-like compounds. The term "lipid-like compound," also referred to as "lipidoid," refers to lipid-like compounds (e.g., amphiphilic compounds with lipid-like physical properties).

[0090] The term " lipid nanoparticle " or " LNP " refers to a particle with at least one nanometer (nm) size (for example, 1 to 1,000nm), which contains one or more types of lipid molecules. LNP provided herein can further contain at least one non-lipid payload molecule (for example, one or more nucleic acid molecules). In some embodiments, LNP comprises a non-lipid payload molecule that is partially or completely encapsulated in lipid shell inside. Especially, in some embodiments, wherein payload is a negatively charged molecule (for example, mRNA encoding viral protein), and the lipid component of LNP comprises at least one cationic lipid. Without being bound by theory, it is expected that cationic lipid can interact with negatively charged payload molecule, and promotes payload to be combined and / or be encapsulated in LNP during LNP formation. Other lipids that can form a part for LNP as provided herein include but are not limited to neutral lipids and charged lipids, such as steroids, polymer-bound lipids and various zwitterionic lipids. In certain embodiments, LNPs according to the present disclosure comprise one or more lipids of series 01, 02, 03, and 04, e.g., one or more lipids of formulas (01-I), (01-II), (02-I), (03-I), and (04-I) (and subformulas thereof) as described herein.

[0091] The term "cationic lipid" refers to a lipid that is positively charged at any pH value or hydrogen ion activity of its environment, or 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). Therefore, the term "cationic lipid" encompasses "permanent cations" and "cationizable". In certain embodiments, the positive charge in the cationic lipid is derived from the presence of a quaternary nitrogen atom. In certain embodiments, the cationic lipid comprises a zwitterionic lipid that is positively charged in the environment of its intended use (e.g., at physiological pH). In certain embodiments, the cationic lipid is one or more series of lipids 01, 02, 03, and 04, such as one or more formulas (01-I), (01-II), (02-I), (03-I), and (04-I) (and subformulas thereof) as described herein. The term "anionic lipid" refers to a lipid that is negatively charged at any pH value or hydrogen ion activity of its environment, or 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). Exemplary anionic lipids include one or more phosphate groups that are negatively charged, eg, at physiological pH.

[0092] The term "polymer-bound lipid" refers to a molecule comprising both a lipid portion and a polymer portion. An example of a polymer-bound lipid is a pegylated lipid (PEG-lipid), wherein the polymer portion comprises polyethylene glycol.

[0093] The term "neutral lipid" encompasses any lipid molecule that exists in an uncharged form or in a neutral zwitterionic form at a selected pH or within a selected pH range. In some embodiments, the selected useful pH or range corresponds to the pH conditions in the environment of the intended lipid use, such as physiological pH. As non-limiting examples, neutral lipids that can be used in conjunction with the present disclosure include, but are not limited to, phosphatidylcholines, 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), 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 hydrogenphosphate (DOCP); sphingomyelin (SM); ceramides; steroids, such as sterols and their derivatives. The neutral lipids provided herein can be synthetic or derived from (isolated or modified from) natural sources or compounds.

[0094] The term "charged lipid" encompasses any lipid molecule present in a positively charged or negatively charged form under a selected pH value or within a selected pH range. In some embodiments, the pH conditions in the environment of the intended lipid use, such as physiological pH, are corresponding to selected pH values ​​or ranges. As non-limiting examples, the neutral lipids that can be used in conjunction with the present disclosure include but are not limited to phosphatidylserine, phosphatidic acid, phosphatidylglycerol, phosphatidylinositol, sterol hemisuccinate, dialkyltrimethylammonium-propane (such as DOTAP, DOTMA), dialkyldimethylaminopropane, ethylphosphocholine, dimethylaminoethanecarbamoylsterol (such as DC-Chol), 1,2-dioleoyl-sn-glycero-3-phosphoric acid-L-serine sodium salt (DOPS-Na), 1,2-dioleoyl-sn-glycero-3-phosphoric acid-(1'-racemic-glycerol) sodium salt (DOPG-Na) and 1,2-dioleoyl-sn-glycero-3-phosphoric acid sodium salt (DOPA-Na). Charged lipids as provided herein can be synthetic or derived from (isolated or modified from) natural sources or compounds.

[0095] As used herein and unless otherwise indicated, the term "alkyl" refers to a saturated straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms. In one embodiment, an alkyl group has, for example, one to twenty-four carbon atoms (C1-C 24 alkyl), four to twenty carbon atoms (C4-C 20 Alkyl), six to sixteen carbon atoms (C6-C 16 alkyl), six to nine carbon atoms (C6-C9 alkyl), one to fifteen carbon atoms (C1-C 15 alkyl), one to twelve carbon atoms (C1-C 12 The term "alkyl" refers to a group consisting of one to eight carbon atoms (C1-C8 alkyl), one to eight carbon atoms (C1-C8 alkyl), or one to six carbon atoms (C1-C6 alkyl) connected to the rest of the molecule by a single bond. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, and the like. Unless otherwise indicated, alkyl groups are optionally substituted.

[0096] As used herein and unless otherwise indicated, the term "alkenyl" refers to a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing one or more carbon-carbon double bonds. As will be appreciated by those of ordinary skill in the art, the term "alkenyl" also encompasses groups having "cis" and "trans" configurations, or groups having "E" and "Z" configurations. In one embodiment, an alkenyl group has, for example, two to twenty-four carbon atoms (C2-C 24 alkenyl), four to twenty carbon atoms (C4-C 20 alkenyl), six to sixteen carbon atoms (C6-C16 alkenyl), six to nine carbon atoms (C6-C9 alkenyl), two to fifteen carbon atoms (C2-C 15 alkenyl), two to twelve carbon atoms (C2-C 12 The term "alkenyl" refers to a group consisting of two to eight carbon atoms (C-C alkenyl), two to eight carbon atoms (C-C alkenyl), or two to six carbon atoms (C-C alkenyl), and is connected to the rest of the molecule by a single bond. Examples of alkenyl groups include, but are not limited to, vinyl, prop-1-enyl, but-1-enyl, pent-1-enyl, pent-1,4-dienyl, etc. Unless otherwise indicated, alkenyl groups are optionally substituted.

[0097] As used herein and unless otherwise indicated, the term "alkynyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing one or more carbon-carbon triple bonds. In one embodiment, an alkynyl group has, for example, two to twenty-four carbon atoms (C2-C 24 Alkynyl), four to twenty carbon atoms (C4-C 20 Alkynyl), six to sixteen carbon atoms (C6-C 16 Alkynyl), six to nine carbon atoms (C6-C9 alkynyl), two to fifteen carbon atoms (C2-C 15 Alkynyl), two to twelve carbon atoms (C2-C 12 Alkynyl is an alkyl radical having two to eight carbon atoms (C2-C8 alkynyl), or two to six carbon atoms (C2-C6 alkynyl) and is connected to the rest of the molecule by a single bond. Examples of alkynyl include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, etc. Unless otherwise indicated, alkynyl is optionally substituted.

[0098] As used herein and unless otherwise indicated, the term "alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain that connects the rest of the molecule to a radical, consisting solely of carbon and hydrogen and being saturated. In one embodiment, an alkylene group has, for example, one to twenty-four carbon atoms (C1-C 24 Alkylene), one to fifteen carbon atoms (C1-C 15 Alkylene), one to twelve carbon atoms (C1-C 12 The alkylene group may be any of a plurality of carbon atoms (C1-C8 alkylene), one to eight carbon atoms (C1-C8 alkylene), one to six carbon atoms (C1-C6 alkylene), two to four carbon atoms (C2-C4 alkylene), one to two carbon atoms (C1-C2 alkylene). Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, n-butylene, and the like. The alkylene chain is connected to the rest of the molecule by a single bond and to the radical group by a single bond. The point of attachment of the alkylene chain to the rest of the molecule and to the radical group may be through one carbon or any two carbon atoms within the chain. Unless otherwise indicated, the alkylene chain is optionally substituted.

[0099] As used herein and unless otherwise indicated, the term "alkenylene" refers to a straight or branched divalent hydrocarbon chain that connects the rest of the molecule to a radical, consisting solely of carbon and hydrogen and containing one or more carbon-carbon double bonds. In one embodiment, an alkenylene group has, for example, two to twenty-four carbon atoms (C2-C 24 alkenyl), two to fifteen carbon atoms (C2-C 15 alkenyl), two to twelve carbon atoms (C2-C 12 Alkenylene is a 1- to 2-carbon group, having two to eight carbon atoms (C-C alkenylene), two to six carbon atoms (C-C alkenylene), or two to four carbon atoms (C-C alkenylene). Examples of alkenylene include, but are not limited to, vinylene, propenylene, n-butenyl, and the like. Alkenylene is attached to the rest of the molecule by a single or double bond and to the radical group by a single or double bond. The points of attachment of the alkenylene to the rest of the molecule and to the radical group may be through one carbon or any two carbon atoms within the chain. Unless otherwise indicated, an alkenylene group is optionally substituted.

[0100] As used herein and unless otherwise indicated, the term "cycloalkyl" refers to a saturated, non-aromatic, monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms. Cycloalkyls may include fused or bridged ring systems. In one embodiment, a cycloalkyl has, for example, 3 to 15 ring carbon atoms (C3-C 15 cycloalkyl), 3 to 10 ring carbon atoms (C3-C 10 Cycloalkyl) or 3 to 8 ring carbon atoms (C3-C8 cycloalkyl). Cycloalkyl is connected to the rest of the molecule by a single bond. Examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic cycloalkyls include, but are not limited to, adamantyl, norbornyl, decahydronaphthyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, and the like. Unless otherwise indicated, cycloalkyl is optionally substituted.

[0101] As used herein and unless otherwise specified, the term "cycloalkylene" is a divalent cycloalkyl group. Unless otherwise specified, a cycloalkylene group is optionally substituted.

[0102] As used herein and unless otherwise indicated, the term "cycloalkenyl" refers to a non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms and containing one or more carbon-carbon double bonds. Cycloalkenyl groups may include fused or bridged ring systems. In one embodiment, a cycloalkenyl group has, for example, 3 to 15 ring carbon atoms (C3-C 15 cycloalkenyl), 3 to 10 ring carbon atoms (C3-C 10The cycloalkenyl group is optionally substituted with 3 to 8 ring carbon atoms (C3-C8 cycloalkenyl). The cycloalkenyl group is connected to the rest of the molecule by a single bond. Examples of monocyclic cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, etc. Unless otherwise indicated, cycloalkenyl groups are optionally substituted.

[0103] As used herein and unless otherwise specified, the term "cycloalkenylene" is a divalent cycloalkenyl group. Unless otherwise specified, a cycloalkenylene group is optionally substituted.

[0104] As used herein and unless otherwise indicated, the term "heterocyclyl" refers to a non-aromatic monocyclic or polycyclic moiety containing one or more (e.g., one, one or two, one to three, or one to four) heteroatoms independently selected from nitrogen, oxygen, phosphorus, and sulfur. The heterocyclyl group may be attached to the main structure at any heteroatom or carbon atom. The heterocyclyl group may be a monocyclic, bicyclic, tricyclic, tetracyclic, or other polycyclic ring system, wherein the polycyclic ring system may be a fused, bridged, or spirocyclic ring system. The heterocyclyl polycyclic ring system may contain one or more heteroatoms in one or more rings. The heterocyclyl group may be saturated or partially unsaturated. Saturated heterocycloalkyl groups may be referred to as "heterocycloalkyl." Partially unsaturated heterocycloalkyl groups may be referred to as "heterocycloalkenyl" when the heterocyclyl group contains at least one double bond, or as "heterocycloalkynyl" when the heterocyclyl group contains at least one triple bond. In one embodiment, the heterocyclyl group 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 (3 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). When appearing herein, a numerical range such as "3 to 18" refers to each integer in the given range; for example, "3 to 18 ring atoms" means that the heterocyclyl group can consist of 3 ring atoms, 4 ring atoms, 5 ring atoms, 6 ring atoms, 7 ring atoms, 8 ring atoms, 9 ring atoms, 10 ring atoms, etc. (up to and including 18 ring atoms). The example of heterocyclic radical includes but is not limited to imidazolyl, imidazolidinyl, oxazolyl, oxazolidinyl, thiazolyl, thiazolidinyl, pyrazolidinyl, pyrazolyl, isoxazolidinyl, isoxazolyl, isothiazolidinyl, isothiazolyl, morpholinyl, pyrrolyl, pyrrolidinyl, furyl, tetrahydrofuranyl, thienyl, pyridyl, piperidyl, quinolyl and isoquinolyl.Unless otherwise stated, heterocyclic radical is optionally substituted.Unless otherwise stated, heterocyclic radical is optionally substituted.

[0105] As used herein, and unless otherwise specified, the term "heterocyclylene" is a divalent heterocyclyl group. Unless otherwise specified, a heterocyclylene group is optionally substituted.

[0106] As used herein and unless otherwise indicated, 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, an aryl group 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 Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthracenyl, phenanthrenyl, pyrenyl, biphenyl, and terphenyl. The term "aryl" also refers to a bicyclic, tricyclic, or other polycyclic hydrocarbon ring, wherein at least one ring is aromatic, and the other rings may be saturated, partially unsaturated, or aromatic, such as dihydronaphthyl, indenyl, dihydroindenyl, or tetrahydronaphthyl (tetrahydronaphthyl / tetralinyl). Unless otherwise indicated, aryl groups are optionally substituted.

[0107] As used herein and unless otherwise specified, the term "arylene" is a divalent aromatic group. Unless otherwise specified, an arylene group is optionally substituted.

[0108] As used herein and unless otherwise indicated, the term "heteroaryl" refers to a monocyclic aromatic group and / or a polycyclic aromatic group containing at least one aromatic ring, wherein at least one aromatic ring contains one or more (e.g., one, one or two, one to three, or one to four) heteroatoms independently selected from O, S, and N. The heteroaryl group can be attached to the main structure at any heteroatom or carbon atom. In certain embodiments, the heteroaryl group has 5 to 20, 5 to 15, or 5 to 10 ring atoms. The term "heteroaryl" also refers to a bicyclic, tricyclic, or other polycyclic ring, wherein at least one ring is an aromatic ring, and the other rings may be saturated, partially unsaturated, or aromatic rings, wherein at least one aromatic ring contains one or more heteroatoms independently selected from O, S, and N. The example of monocyclic heteroaryl includes but is not limited to pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl. The example of bicyclic heteroaryl includes but is not limited to indolyl, benzothiazolyl, benzoxazolyl, benzothienyl, quinolyl, tetrahydroisoquinolyl, isoquinolyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, isobenzofuranyl, chromone radical, coumarin base, cinnolinyl, quinoxalinyl, indazolyl, purinyl, pyrrolopyridinyl, furopyridinyl, thienopyridinyl, dihydroisoindolyl and tetrahydroquinolyl. Examples of tricyclic heteroaryl groups include, but are not limited to, carbazolyl, benzindolyl, phenanthrolinyl, acridinyl, phenanthridinyl, and xanthenyl.Unless otherwise specified, heteroaryl groups are optionally substituted.

[0109] As used herein and unless otherwise specified, the term "heteroarylene" is a divalent heteroaryl group. Unless otherwise specified, a heteroarylene group is optionally substituted.

[0110] When a group described herein is referred to as "substituted," it may be substituted with one or more suitable substituents. Illustrative examples of substituents include, but are not limited to, those found in the exemplary compounds and embodiments provided herein, as well as: halogen atoms, such as F, Cl, 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 independently H, C1-C 15 alkyl or cycloalkyl, and x is 0, 1 or 2. In some embodiments, the substituent is C1-C 12 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').

[0111] As used herein and unless otherwise indicated, the term "optional" or "optionally" (e.g., optionally substituted) means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted alkyl" means that the alkyl group may or may not be substituted, and that the description includes both substituted alkyl groups and alkyl groups without substitution.

[0112] As used herein and unless otherwise indicated, the term "prodrug" of a biologically active compound refers to a compound that can be converted into a biologically active compound under physiological conditions or by solvent decomposition. In one embodiment, the term "prodrug" refers to a pharmaceutically acceptable metabolic precursor of a biologically active compound. The prodrug may be inactive when administered to a subject in need, but is converted into a biologically active compound in vivo. Prodrugs are generally rapidly converted in vivo to produce the parent biologically active compound, for example, by hydrolysis in the blood. Prodrug compounds generally provide advantages of solubility, tissue compatibility, or delayed release in mammalian organisms (see Bundgard, H., Design of Prodrugs (1985), pp. 7-9, pp. 21-24 (Elsevier, Amsterdam)). Discussions on prodrugs are provided in Higuchi, T. et al., ACS Symposium Series, Vol. 14; and Bioreversible Carriers in Drug Design, edited by Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

[0113] In one embodiment, the term "prodrug" is also intended to include any covalently bonded carriers that release the active compound in vivo when such prodrugs are administered to a mammalian subject. Prodrugs of a compound can be prepared by modifying the functional groups present in the compound in such a way that the modifications can be cleaved during routine manipulation or in vivo to yield the parent compound. Prodrugs include compounds in which a hydroxyl, amino, or sulfhydryl group is bonded to any group that, when the prodrug of the compound is administered to a mammalian subject, cleaves to form a free hydroxyl, free amino, or free sulfhydryl group, respectively.

[0114] Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of alcohol functional groups or amide derivatives of amine functional groups in the compounds provided herein.

[0115] As used herein, and unless otherwise indicated, the term "pharmaceutically acceptable salt" includes both acid addition salts and base addition salts.

[0116] Examples of pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; and organic acids such as, but 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, carbonic acid, cinnamic acid, citric acid, cyclamic acid, benzo ... acid), dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxoglutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic 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, undecylenic acid, and the like.

[0117] Examples of pharmaceutically acceptable base addition salts include, but are not limited to, salts prepared by adding an inorganic base or an organic base to a free acid compound. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. In one embodiment, the inorganic salt is an ammonium salt, a sodium salt, a potassium salt, a calcium salt, and a magnesium salt. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, denanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, phenethylbenzylamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. In one embodiment, the organic base is isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0118] The compounds provided herein may contain one or more asymmetric centers and thus may produce enantiomers, diastereomers, and other stereoisomeric forms, which may be defined as (R)- or (S)- or as (D)- or (L)- for amino acids based on absolute stereochemistry. Unless otherwise indicated, the compounds provided herein are intended to 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 may be prepared using chiral synthons or chiral reagents, or 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 resolving racemates (or racemates of salts or derivatives) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other geometric asymmetric centers, unless otherwise indicated, the compounds are intended to include E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included.

[0119] As used herein and unless otherwise indicated, the term "isomer" refers to different compounds with the same molecular formula. "Stereoisomers" are isomers that differ only in the arrangement of their atoms in space. "Atropisomers" are stereoisomers resulting from hindered rotation about a single bond. "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A mixture of a pair of enantiomers in any ratio is referred to as a "racemic" mixture. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other.

[0120] "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 E or Z isomers. In other embodiments, the compounds described herein are mixtures of E and Z isomers.

[0121] "Tautomers" refer to isomeric forms of a compound that are in equilibrium with each other. The concentrations of the isomeric forms will depend on the environment in which the compound is located, and may vary depending on, for example, whether the compound is a solid or in an organic or aqueous solution.

[0122] It should also be noted that the compounds described herein may contain unnatural proportions of atomic isotopes at one or more atoms. For example, the compounds may be substituted with radioactive isotopes such as tritium ( 3 H), iodine-125 ( 125 I), sulfur-35( 35 S) or carbon-14 ( 14 C) is radiolabeled, or may be isotopically enriched, such as deuterium ( 2 H), carbon-13 (13 C) or nitrogen-15( 15 N). As used herein, an "isotopologue" is an isotopically enriched compound. The term "isotopically enriched" refers to an atom whose isotopic composition is different from the natural isotopic composition of the atom. "Isotopically enriched" may also refer to a compound containing at least one atom whose isotopic composition is different from the natural isotopic composition of the atom. The term "isotopic composition" refers to the amount of each isotope present in a given atom. Radiolabeled and isotopically enriched compounds can be used as therapeutic agents, such as cancer therapeutics; research reagents, such as binding assay reagents; and diagnostic agents, such as in vivo imaging agents. All isotopic variants of the compounds described herein, whether radioactive or not, are intended to be encompassed within the scope of the embodiments provided herein. In some embodiments, isotopologues of the compounds described herein are provided, for example, isotopologues that are enriched in deuterium, carbon-13, and / or nitrogen-15. As used herein, "deuterated" refers to a compound in which at least one hydrogen (H) has been replaced by deuterium (as D or 2 H represents) substitution, that is, the compound is enriched in deuterium at at least one position.

[0123] It should be noted that if there is a discrepancy between a depicted structure and the name of the structure, the depicted structure shall control.

[0124] As used herein and unless otherwise indicated, the term "pharmaceutically acceptable carrier, diluent or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the U.S. Food and Drug Administration as acceptable for use in humans or veterinary animals.

[0125] The term "composition" is intended to encompass a product containing specified ingredients (eg, mRNA molecules provided herein), optionally in specified amounts.

[0126] As used interchangeably herein, the terms "polynucleotide" or "nucleic acid" refer to nucleotide polymers of any length, and include, for example, DNA and RNA. Nucleotides may be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases and / or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthetic reaction. Polynucleotides may comprise modified nucleotides, such as methylated nucleotides and their analogs. Nucleic acids may be in single-stranded or double-stranded form. As used herein and unless otherwise indicated, "nucleic acid" also includes nucleic acid mimics, such as locked nucleic acids (LNA), peptide nucleic acids (PNA), and morpholino nucleic acids. As used herein, "oligonucleotide" refers to a short synthetic polynucleotide, generally but not necessarily less than about 200 nucleotides in length. The terms "oligonucleotide" and "polynucleotide" are not mutually exclusive. The above description of polynucleotides is equally and fully applicable to oligonucleotides. Unless otherwise indicated, the left-hand end of any single-stranded polynucleotide sequence disclosed herein is the 5' end; the left-hand direction of a double-stranded polynucleotide sequence is referred to as the 5' direction. The direction of 5' to 3' addition of the nascent RNA transcript is called the transcription direction; the sequence region on the DNA chain that has the same sequence as the RNA transcript and is located 5' relative to the 5' end of the RNA transcript is called the "upstream sequence"; the sequence region on the DNA chain that has the same sequence as the RNA transcript and is located 3' relative to the 3' end of the RNA transcript is called the "downstream sequence".

[0127] As used herein, the term "non-naturally occurring" when used with reference to nucleic acid molecules as described herein is intended to mean that the nucleic acid molecules do not exist in nature. Non-naturally occurring nucleic acids encoding viral peptides or proteins contain at least one genetic alteration or chemical modification that is not typically present in naturally occurring strains of the virus (including wild-type strains of the virus). Genetic alteration includes, for example, modification of an expressible nucleic acid sequence that introduces a heterologous peptide or polypeptide encoding the virus, other nucleic acid additions, nucleic acid deletions, nucleic acid substitutions, and / or other functional disruptions to the viral genetic material. Such modifications include, for example, modification of the coding region and functional fragments thereof of heterologous, homologous, or heterologous and homologous polypeptides of the viral species. Additional modifications include, for example, modification of non-coding regulatory regions, wherein the modifications alter the expression of a gene or operon. Additional modifications also include, for example, incorporating the nucleic acid sequence into a vector such as a plasmid or artificial chromosome. Chemical modifications include, for example, one or more functional nucleotide analogs as described herein.

[0128] " isolated nucleic acid " refers to nucleic acid, for example RNA, DNA or mixed nucleic acid, that is basically separated from other genomic DNA sequences and proteins or complexes (such as ribosomes and polymerases) that naturally follow the native sequence." isolated " nucleic acid molecule is a nucleic acid molecule separated from other nucleic acid molecules that are present in the natural origin of the nucleic acid molecule. In addition, when manufactured by recombinant technology, the nucleic acid molecule of " isolation ", such as mRNA molecule, can be basically free of other cell materials or culture medium, or when chemically synthesized, it can be basically free of chemical precursors or other chemicals. In specific embodiments, one or more nucleic acid molecules of encoding antigens as described herein are isolated or purified. The term includes the nucleic acid sequence removed from its naturally occurring environment, and includes recombinant or cloned DNA or RNA isolates and chemically synthesized analogs or analogs biosynthesized by heterologous systems. Substantially pure molecule can include the isolated form of molecule.

[0129] The term "coding nucleic acid" or its grammatical equivalents when used to refer to a nucleic acid molecule includes: (a) a nucleic acid molecule that can be transcribed to produce mRNA and then translated into peptides and / or polypeptides when in its natural state or manipulated by methods well known to those skilled in the art; and (b) the mRNA molecule itself. The antisense strand is the complementary sequence of such a nucleic acid molecule, and the coding sequence can be inferred therefrom. The term "coding region" refers to the portion of a coding nucleic acid sequence that is translated into a peptide or polypeptide. The term "untranslated region" or "UTR" refers to the portion of a coding nucleic acid that is not translated into a peptide or polypeptide. Depending on the orientation of the UTR relative to the coding region of the nucleic acid molecule, the UTR is referred to as a 5'-UTR if it is located at the 5' end of the coding region, and as a 3'-UTR if it is located at the 3' end of the coding region.

[0130] As used herein, the term "mRNA" refers to a messenger RNA molecule comprising one or more open reading frames (ORFs), which can be translated by a cell or organism having the 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 also comprises one or more untranslated regions (UTRs).

[0131] In certain embodiments, the mRNA is a monocistronic mRNA comprising only one ORF. In certain embodiments, the monocistronic mRNA encodes a peptide or protein comprising 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 polycistronic mRNA comprising two or more ORFs. In certain embodiments, the polycistronic mRNA encodes two or more peptides or proteins that may be identical or different from each other. In certain embodiments, each peptide or protein encoded by the polycistronic mRNA comprises at least one epitope of a selected antigen. In certain embodiments, the different peptides or proteins encoded by the polycistronic mRNA each comprise at least one epitope of a different antigen. In any of the embodiments described herein, the at least one epitope may be at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 epitopes of an antigen.

[0132] The term "nucleobase" encompasses purines and pyrimidines, including the natural compounds adenine, thymine, guanine, cytosine, uracil, inosine, and natural or synthetic analogs or derivatives thereof.

[0133] As used herein, the term "functional nucleotide analogue" refers to a modified version of a classical nucleotide A, G, C, U, or T that (a) retains the base pairing properties of the corresponding classical nucleotide and (b) contains at least one chemical modification of the (i) nucleobase, (ii) sugar group, (iii) phosphate group, or (iv) any combination of (i) to (iii) of the corresponding natural nucleotide. As used herein, base pairing encompasses not only the classical Watson-Crick adenine-thymine, adenine-uracil, or guanine-cytosine base pairs, but also base pairs formed between a classical nucleotide and a functional nucleotide analogue, or between a pair of functional nucleotide analogues, wherein the arrangement of hydrogen bond donors and hydrogen bond acceptors allows hydrogen bonds to form between the modified nucleobase and the classical nucleobase, or between two complementary modified nucleobase structures. For example, a functional analogue of guanosine (G) retains the ability to base pair with cytosine (C) or a functional analogue of cytosine. An example of such non-classical base pairing is base pairing between a modified nucleotide inosine and adenine, cytosine, or uracil. As described herein, functional nucleotide analogs can be naturally occurring or non-naturally occurring. Thus, nucleic acid molecules containing functional nucleotide analogs can have at least one modified nucleobase, sugar group, and / or internucleoside linkage. Exemplary chemical modifications to nucleobases, sugar groups, or internucleoside linkages of nucleic acid molecules are provided herein.

[0134] As used herein, the terms "translational enhancer element," "TEE," and "translational enhancer" refer to regions in nucleic acid molecules that promote translation of a nucleic acid coding sequence into a protein or peptide product, such as via cap-dependent or cap-independent translation. TEEs are typically located in the UTR region of a nucleic acid molecule (e.g., mRNA) and enhance the translation level of a coding sequence 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, August 2013; 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 (Pánek et al. Nucleic Acids Research, Vol. 41, No. 16, Sept. 1, 2013, pp. 7625-7634).

[0135] 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 an unpaired loop. The resulting structure is called a stem-loop structure, hairpin, or hairpin loop, which is a secondary structure found in many RNA molecules.

[0136] As used herein, the term "peptide" refers to a polymer containing from two to fifty (2-50) amino acid residues linked by one or more covalent peptide bonds. 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-naturally occurring amino acids (e.g., amino acid analogs or non-natural amino acids).

[0137] The terms "polypeptide" and "protein" are used interchangeably herein to refer to a polymer having more than fifty (50) amino acid residues linked by covalent peptide bonds. That is, a description of a polypeptide equally applies to a description of a protein, and vice versa. The terms apply to naturally occurring amino acid polymers as well as amino acid polymers in which one or more amino acid residues is a non-naturally occurring amino acid (e.g., an amino acid analog). As used herein, the terms encompass amino acid chains of any length, including full-length proteins (e.g., antigens).

[0138] In the case of a peptide or polypeptide, 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 introducing an amino acid residue substitution, deletion, or addition. 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 attaching any type of molecule to a polypeptide. For example, but not limited to, a viral peptide or protein or a fragment of a viral peptide or protein can be chemically modified, for example, by glycosylation, acetylation, pegylation, phosphorylation, amidation, by derivatization of known protecting groups / blocking groups, proteolytic cleavage, chemical cleavage, formulation, tunicamycin metabolic synthesis, connection to a cell ligand or other protein, etc. Derivatives are modified in a manner different from naturally occurring or starting peptides or polypeptides in terms of the type or position of the attached molecule. Derivatives also include the absence of one or more naturally occurring chemical groups on a viral peptide or protein. In addition, the derivatives of a viral peptide or protein or a fragment of a viral peptide or protein can contain one or more non-classical amino acids. In specific embodiments, the derivative is a functional derivative of the native or unmodified peptide or polypeptide from which the derivative is derived.

[0139] 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 the derivative is derived. 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 RNA or package the viral genome.

[0140] The term "identity" refers to the relationship between the sequences of two or more polypeptide molecules or two or more nucleic acid molecules, as determined by aligning and comparing the sequences. "Percentage (%) of amino acid sequence identity" relative to a reference polypeptide sequence is defined as the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence after aligning the sequences and, if necessary, introducing gaps (to achieve maximum sequence identity) and not considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining percentage of amino acid sequence identity can be achieved in a variety of ways within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or MEGALIGN (DNAStar, Inc.) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm required for achieving maximum alignment over the full length of the compared sequences.

[0141] "Modification" of an amino acid residue / position refers to a change in the primary amino acid sequence as compared to the starting amino acid sequence, wherein the change is caused by a sequence change involving the amino acid residue / position. For example, typical modifications include replacing a residue with another amino acid (e.g., conservative or non-conservative substitutions), inserting one or more (e.g., typically less than 5, 4, or 3) amino acids immediately adjacent to the residue / position, and / or deleting the residue / position.

[0142] In the case of a peptide or polypeptide, as used herein, the term "fragment" refers to a peptide or polypeptide that comprises an amino acid sequence that is less than the full length. Such fragments may, for example, come from truncation of the amino terminus, truncation of the carboxyl terminus, and / or internal deletions of residues in the amino acid sequence. Fragments may, for example, be produced by alternative RNA splicing or by in vivo protease activity. In certain embodiments, a fragment refers to at least 5 consecutive amino acid residues, at least 10 consecutive amino acid residues, at least 15 consecutive amino acid residues, at least 20 consecutive amino acid residues, at least 25 consecutive amino acid residues, at least 30 consecutive amino acid residues, at least 40 consecutive amino acid residues, at least 50 consecutive amino acid residues, at least 60 consecutive amino acid residues, at least 70 consecutive amino acid residues, at least 80 consecutive amino acid residues, at least 90 consecutive amino acid residues, at least 100 consecutive amino acid residues, or at least 110 consecutive amino acid residues. In some embodiments, a fragment of a polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues, 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 a specific embodiment, a fragment of a polypeptide retains at least one, at least two, at least three or more functions of the polypeptide.

[0143] As used herein in the context of a peptide or polypeptide (e.g., a protein), the term "immunogenic fragment" refers to a fragment of a peptide or polypeptide that retains the ability of the peptide or polypeptide to elicit an immune response (including an innate immune response and / or an adaptive immune response) when in contact with a mammalian immune system. In some embodiments, an immunogenic fragment of a peptide or polypeptide can be an epitope.

[0144] The term "antigen" refers to a substance that can be recognized by a subject's immune system (including the adaptive immune system) and that can trigger an immune response (including an antigen-specific immune response) after contacting the subject with the antigen. In certain embodiments, the antigen is a protein associated with a diseased cell (such as a cell infected by a pathogen) or a neoplastic cell (e.g., a tumor-associated antigen (TAA)).

[0145] An "epitope" is a site on the surface of an antigen molecule that binds to a single antibody molecule, such as a localized region on the surface of an antigen that is capable of binding to one or more antigen-binding regions of an antibody and that has antigenic or immunogenic activity capable of eliciting an immune response in an animal, such as a mammal (e.g., a human). An epitope with immunogenic activity is a portion of a polypeptide that elicits an antibody response in an animal. An epitope with antigenic activity is a portion of a polypeptide to which an antibody binds, as determined by any method known in the art, including, for example, by immunoassay. An antigenic epitope is not necessarily immunogenic. Epitopes are typically composed of chemically active surface groups of a molecule, such as amino acids or sugar side chains, and have specific three-dimensional structural characteristics as well as specific charge characteristics. Antibody epitopes can be linear or conformational. Linear epitopes are formed by contiguous sequences of amino acids in a protein. Conformational 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 three-dimensional structure of a protein assumes an altered conformation, such as after activation or binding by another protein or ligand. In certain embodiments, an epitope is a three-dimensional surface feature of a polypeptide. In other embodiments, an epitope is a linear feature of a polypeptide. Typically, an antigen has several or many different epitopes and can react with many different antibodies.

[0146] The term "heterologous" refers to an entity not found in nature related to (e.g., encoded by and / or expressed by) a naturally occurring VZV. The term "homologous" refers to an entity found in nature related to (e.g., encoded by and / or expressed by) a naturally occurring VZV.

[0147] As used herein, the term "genetic vaccine" refers to a therapeutic or preventive 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). Administering a vaccine to a subject ("vaccination") allows the production of encoded peptides or proteins, thereby triggering an immune response against the target disease in the subject. In certain embodiments, the immune response includes an adaptive immune response, such as the production of antibodies against the encoded antigen, and / or the activation and proliferation of immune cells capable of specifically eliminating diseased cells expressing the antigen. In certain embodiments, the immune response also includes an innate immune response. According to the present disclosure, the vaccine can be administered to a subject before or after the onset of clinical symptoms of the target disease. In some embodiments, vaccination of healthy or asymptomatic subjects makes the vaccinated subject immune or less sensitive to the development of the target disease. In some embodiments, vaccination of subjects showing symptoms of the disease improves the disease condition of the vaccinated subject or treats the disease.

[0148] The term "vector" refers to a material for carrying or comprising a nucleic acid sequence, including, for example, a nucleic acid sequence encoding a viral peptide or protein as described herein, so that the nucleic acid sequence is introduced into a host cell, or used as a transcription template to carry out in vitro transcription reaction in a cell-free system to produce mRNA. Suitable vectors for use include, for example, expression vectors, plasmids, phage vectors, viral vectors, episomes, and artificial chromosomes, which may include a selection sequence or a marker operable for stable integration into the chromosome of a host cell. In addition, the vector may include one or more selective marker genes and appropriate transcription or translation control sequences. For example, the selective marker gene that may be included provides resistance to antibiotics or toxins, supplements nutritional deficiencies, or provides key nutrients that are not in the culture medium. Transcription or translation control sequences may include constitutive and inducible promoters, transcription enhancers, transcription terminators, etc. as known in the art. When two or more nucleic acid molecules (for example, nucleic acid molecules encoding two or more different viral peptides or proteins) are co-transcribed or co-translated, the two nucleic acid molecules may be inserted, for example, into the same expression vector or into a separate expression vector. For single vector transcription and / or translation, the encoding nucleic acid can be operably linked to a common transcription or translation control sequence, or to different transcription or translation control sequences, such as an inducible promoter and a constitutive promoter. Methods well known in the art can be used to confirm the introduction of nucleic acid molecules into host cells. Such methods include, for example, nucleic acid analysis, such as Northern blot or polymerase chain reaction (PCR) amplification of mRNA; immunoblotting for expression of gene products; or other suitable analytical methods for testing the expression of the introduced nucleic acid sequence or its corresponding gene product. It will be understood by those skilled in the art that nucleic acid molecules are expressed in sufficient amounts to produce the desired product (e.g., mRNA transcripts of nucleic acids as described herein), and it will be further understood that expression levels can be optimized using methods well known in the art to obtain sufficient expression.

[0149] The terms "innate immune response" and "innate immunity" are recognized in the art and refer to nonspecific defense mechanisms that the body's immune system initiates when it recognizes pathogen-associated molecular patterns, which involve different forms of cellular activity, including cytokine production and cell death through various pathways. As used herein, the 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 in some cases, induction of apoptosis. Activation of innate immunity can be detected using methods known in the art, such as measuring (NF)-κB activation.

[0150] 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 when it recognizes a specific antigen, including humoral and cell-mediated responses. As used herein, an adaptive immune response includes a cellular response triggered and / or enhanced by a vaccine composition, such as the genetic composition described herein. In some embodiments, the vaccine composition comprises an antigen that is a target of an antigen-specific adaptive immune response. In other embodiments, the vaccine composition, upon administration, allows the production of an antigen in the immunized subject that is a target of an antigen-specific adaptive immune response. Activation of the adaptive immune response can be detected using methods known in the art, such as measuring the production of antigen-specific antibodies or the level of antigen-specific cell-mediated cytotoxicity.

[0151] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted 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 target cells bearing antigens and subsequently kill the target cells with cytotoxins. Antibodies "arm" the cytotoxic cells and are absolutely required for this killing. NK cells (the primary cells for mediating ADCC) express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is known (see, e.g., Ravetch and Kinet, 1991, Annu. Rev. Immunol. 9:457-92). To assess the ADCC activity of a target molecule, an in vitro ADCC assay can be performed (see, e.g., 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 can be assessed in vivo, for example, 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 can be selected for use.

[0152] "Antibody-dependent cellular phagocytosis" or "ADCP" refers to the destruction of target cells via monocyte- or macrophage-mediated phagocytosis when immunoglobulins bind to Fc receptors (FcRs) present on certain phagocytes (e.g., neutrophils, monocytes, and macrophages) so that these phagocytes can specifically bind to target cells carrying antigens and subsequently kill target cells. In order to assess the ADCP activity of a target molecule, an in vitro ADCP assay (see, e.g., Bracher et al., 2007, J. Immunol. Methods 323: 160-71) can be performed. Useful phagocytes for such assays include peripheral blood mononuclear cells (PBMCs), purified monocytes from PBMCs, or U937 cells differentiated into mononuclear types. Alternatively or in addition, the ADCP activity of a target molecule can be evaluated in vivo, for example, in an animal model (see, e.g., Wallace et al., 2001, J. Immunol. Methods 248: 167-82). Antibodies with little or no ADCP activity can be selected for use.

[0153] "Fc receptor" or "FcR" describes a receptor that binds to the Fc region of an antibody. An exemplary FcR is a native sequence human FcR. Additionally, an exemplary FcR is a receptor that binds to IgG antibodies (e.g., a gamma receptor), and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced ​​forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains (see, e.g., 1997, Annu. Rev. Immunol. 15: 203-34). Various FcRs are known (see, e.g., Ravetch and Kinet, 1991, Annu. Rev. 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). The term "FcR" herein encompasses other FcRs, including those to be identified in the future. The term also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgG to the fetus (see, e.g., 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 decreased binding to FcRs have been described (see, eg, WO 2000 / 42072; US Patent Nos. 7,183,387; 7,332,581; and 7,335,742; Shields et al., 2001, J. Biol. Chem. 9(2):6591-604).

[0154] "Complement-dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by binding of the first component of the complement system (C1q) to an antibody (of the appropriate subclass) bound to its cognate antigen. To assess complement activation, a CDC assay can be performed (see, e.g., Gazzano-Santoro et al., 1996, J. Immunol. Methods 202:163). Polypeptide variants with altered Fc region amino acid sequences (polypeptides with variant Fc regions) and increased or decreased C1q binding capacity have been described (see, e.g., U.S. Patent No. 6,194,551; WO 1999 / 51642; Idusogie et al., 2000, J. Immunol. 164:4178-84). Antibodies with little or no CDC activity can be selected for use.

[0155] The term "antibody" is intended to include polypeptide products of B cells within the class of immunoglobulin polypeptides, which are capable of binding to specific molecular antigens and consisting of two pairs of identical polypeptide chains, wherein each pair has a heavy chain (about 50-70 kDa) and a light chain (about 25 kDa), each amino terminal portion of each chain comprises a variable region of about 100 to about 130 or more amino acids, and each carboxyl terminal portion of each chain comprises a constant region. See, for example, Antibody Engineering (Borrebaeck ed., 2nd ed., 1995); and Kuby, Immunology (3rd ed., 1997). In specific embodiments, specific molecular antigens can be bound by antibodies provided herein, including polypeptides, fragments thereof, or epitopes. Antibodies also include, but are not limited to, synthetic antibodies, recombinantly produced antibodies, camelized antibodies, intracellular antibodies, anti-idiotypic (anti-Id) antibodies, and functional fragments of any of the above, which refers to a portion of the antibody heavy or light chain polypeptide that retains some or all of the binding activity of the antibody as the source of the fragment. 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-linked Fv (dsFv), Fd fragments, Fv fragments, diabodies, triabodies, tetrabodies and minibodies. In particular, the antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, such as antigen binding domains or molecules containing 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; Plückthun and Skerra, 1989, Meth. Enzymol. 178: 497-515; and Day, Advanced Immunochemistry (2nd ed., 1990). The antibodies provided herein can be of any class (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) of immunoglobulin molecules.

[0156] The term "administer" or "administration" refers to the act of injecting or otherwise physically delivering a substance (e.g., a lipid nanoparticle composition described herein) present in vitro into a patient's body, such as by transmucosal, intradermal, intravenous, intramuscular delivery, and / or any other physical delivery method described herein or known in the art. When treating a disease, condition, illness, or symptom thereof, the administration of the substance is typically performed after the onset of the disease, condition, illness, or symptom thereof. When preventing a disease, condition, illness, or symptom thereof, the administration of the substance is typically performed before the onset of the disease, condition, illness, or symptom thereof.

[0157] "Long-term" administration, as opposed to acute mode, refers to administration of one or more agents in a continuous mode (e.g., for a period of time, such as days, weeks, months, or years), thereby maintaining the initial therapeutic effect (activity) over an extended period of time. "Intermittent" administration refers to treatment that is not continuous without interruption, but rather is cyclical in nature.

[0158] As used herein, the term "targeted delivery" or verb form "targeting" refers to a process in which the delivered agent (such as the therapeutic payload molecule in the lipid nanoparticle composition as described herein) is promoted to reach a specific organ, tissue, cell and / or intracellular compartment (referred to as the target location) compared to delivery to any other organ, tissue, cell or intracellular compartment (referred to as a non-target location). Targeted delivery can be detected using methods known in the art, such as by comparing the concentration of the delivered agent in the target cell population with the concentration of the delivered agent at the non-target cell population after systemic administration. In certain embodiments, targeted delivery makes the concentration at the target location at least 2 times higher than the concentration at the non-target location.

[0159] An "effective amount" is generally an amount sufficient to reduce the severity and / or frequency of symptoms; eliminate symptoms and / or underlying causes; prevent the occurrence of symptoms and / or their underlying causes; and / or ameliorate or remedy damage caused by or associated with a disease, condition, or disorder, including, for example, infection and neoplasia. In some embodiments, an effective amount is a therapeutically effective amount or a prophylactically effective amount.

[0160] As used herein, the term "therapeutically effective amount" refers to an amount of an agent (e.g., a vaccine composition) sufficient to reduce and / or ameliorate the severity and / or duration of a given disease, condition, or disorder, and / or its associated symptoms (e.g., an infectious disease, such as an infectious disease caused by a viral infection, or a neoplastic disease, such as cancer). A "therapeutically effective amount" of a substance / molecule / agent disclosed herein (e.g., a lipid nanoparticle composition described herein) may vary depending on a number of factors, such as the disease state, age, sex, and weight of the individual, and the ability of the substance / molecule / agent to elicit a desired response in the individual. A therapeutically effective amount comprises an amount in which any toxic or deleterious effects of the substance / molecule / agent are outweighed by the therapeutically beneficial effects. In certain embodiments, the term "therapeutically effective amount" refers to an amount of a lipid nanoparticle composition as described herein or a therapeutic or prophylactic agent (e.g., a therapeutic mRNA) contained therein that is effective to "treat" a disease, condition, or disorder in a subject or mammal.

[0161] A "prophylactically effective amount" is an amount of an agent or pharmaceutical composition (e.g., a vaccine composition) that, when administered to a subject, will have the intended prophylactic effect, e.g., prevent a disease, disorder, condition, or associated symptoms (e.g., an infectious disease, such as an infectious disease caused by a viral infection, or a neoplastic disease, such as cancer), delay its onset (or recurrence), or reduce the likelihood of its onset (or recurrence). In certain embodiments, the term "prophylactically effective amount" refers to an amount of a lipid nanoparticle composition described herein, or a prophylactic agent contained therein (e.g., a nucleic acid, such as, for example, a nucleic acid described in Section 5.4 or 6, including mRNA), that is effective to "prevent" a disease, disorder, or condition in a subject or mammal. Typically, but not necessarily, a prophylactic effective amount may be less than a therapeutically effective amount because a prophylactic dose is administered to a subject prior to or at an early stage of a disease, disorder, or condition. A full therapeutic or prophylactic effect may not necessarily occur with the administration of a single dose, but may only occur after a series of doses. Thus, a therapeutically or prophylactically effective amount may be administered in one or more administrations.

[0162] The terms "prevent," "preventing," and "prevention" refer to reducing the likelihood of onset (or recurrence) of a disease, disorder, condition, or associated symptoms, such as an infectious disease, such as caused by a viral infection, or a neoplastic disease, such as cancer.

[0163] The terms "manage," "managing," and "management" refer to the beneficial effects that a subject obtains from a therapy (e.g., a prophylactic or therapeutic agent) that does not result in a cure of the disease. In certain embodiments, one or more therapies (e.g., a prophylactic or therapeutic agent, such as a lipid nanoparticle composition described herein) are administered to a subject to "manage" an infectious or neoplastic disease, or one or more symptoms thereof, thereby preventing progression or worsening of the disease.

[0164] The term "prophylactic agent" refers to any agent that can completely or partially inhibit the development, recurrence, onset or spread of a disease and / or symptoms associated therewith in a subject. In some embodiments, the prophylactic agent comprises or consists of a nucleic acid as described herein (e.g., a nucleic acid described in Section 5.4 or Section 6). In some embodiments, the prophylactic agent comprises or consists of a protein as described herein (e.g., a protein described in Section 5.3). In some embodiments, the prophylactic agent comprises or consists of a vector comprising a nucleic acid as described herein.

[0165] The term "therapeutic agent" refers to any agent that can be used to treat, prevent, or alleviate a disease, condition, or illness, including any agent that can be used to treat, prevent, or alleviate one or more symptoms of a disease, condition, or illness and / or symptoms associated therewith. In some embodiments, the therapeutic agent is a nucleic acid as described herein (e.g., a nucleic acid described in Section 5.4 or Section 6). In some embodiments, the therapeutic agent is a protein as described herein (e.g., a protein described in Section 5.3). In some embodiments, the prophylactic agent comprises or consists of a vector comprising a nucleic acid as described herein.

[0166] The term "therapy" refers to any regimen, method, and / or agent that can be used to prevent, control, treat, and / or improve a disease, condition, or illness. In certain embodiments, the term "therapies" or "therapy" refers to biological therapies, supportive therapies, and / or other therapies known to those skilled in the art, such as medical personnel, that can be used to prevent, control, treat, and / or improve a disease, condition, or illness.

[0167] As used herein, a "prophylactically effective serum titer" is a serum titer of antibodies in a subject (e.g., a human) that completely or partially inhibits the development, recurrence, onset, or spread of a disease, disorder, or condition and / or symptoms associated therewith in the subject.

[0168] In certain embodiments, a "therapeutically effective serum titer" is a serum titer of antibodies 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.

[0169] The term "serum titer" refers to the average serum titer in a subject from multiple samples (e.g., at multiple time points) or in a population of at least 10, at least 20, at least 40 subjects, up to about 100, 1000 or more subjects.

[0170] The term "side effect" encompasses unwanted and / or adverse effects of a therapy (such as a preventive or therapeutic agent). An unwanted effect is not necessarily adverse. The adverse effects of a therapy (such as a preventive or therapeutic agent) may be harmful, uncomfortable, or risky. Examples of side effects include diarrhea, cough, gastroenteritis, wheezing, nausea, vomiting, anorexia, abdominal cramps, fever, pain, weight loss, dehydration, alopecia, dyspnea, insomnia, dizziness, mucositis, nerve and muscle effects, fatigue, dry mouth, loss of appetite, rash or swelling at the site of administration, flu-like symptoms such as fever, chills, and fatigue, digestive tract problems, and allergic reactions. Other undesirable effects experienced by patients are numerous and known in the art. Many effects are described in Physician's Desk Reference (68th edition, 2014).

[0171] The terms "subject" and "patient" are used interchangeably. As used herein, in certain embodiments, the subject is a mammal, such as a non-primate (e.g., a cow, pig, horse, cat, dog, rat, etc.) or a primate (e.g., a monkey and a human). In specific embodiments, the subject is a human. In one embodiment, the subject is a mammal (e.g., a human) suffering from an infectious disease or a neoplastic disease. In another embodiment, the subject is a mammal (e.g., a human) at risk of developing an infectious disease or a neoplastic disease.

[0172] The term "elderly" refers to a person over the age of 65. The term "human adult" refers to a person over the age of 18. The term "human child" refers to a person between the ages of 1 and 18. The term "human toddler" refers to a person between the ages of 1 and 3. The term "human infant" refers to a person between the ages of newborn and 1 year old.

[0173] The term "detectable probe" refers to a composition that provides a detectable signal. The term includes, but is not limited to, any fluorophore, chromophore, radiolabel, enzyme, antibody, or antibody fragment that provides a detectable signal through activity.

[0174] The term "detectable agent" refers to a substance that can be used to determine the presence of a desired molecule, such as an antigen encoded by an mRNA molecule described herein, in a sample or subject. A detectable agent can be a substance that can be visually detected or can be determined and / or measured (e.g., by quantitation) in other ways.

[0175] By "substantially all" is meant 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%.

[0176] As used herein and unless otherwise indicated, the term "about" or "approximately" means an acceptable error for a particular value determined by one of ordinary skill in the art, which depends in part on the manner in which the value is measured or determined. In certain embodiments, the term "about" or "approximately" means within 1, 2, 3, or 4 standard deviations. In certain embodiments, the terms "about" and "approximately" mean within 20%, within 15%, within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3%, within 2%, within 1%, within 0.5%, within 0.05%, or lower of a given value or range.

[0177] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0178] All publications, patent applications, accession numbers, and other references cited in this specification are incorporated herein by reference in their entirety, to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to preempt such publications by virtue of prior invention. In addition, the publication dates provided may differ from the actual publication dates, which may require independent confirmation.

[0179] A number of embodiments of the present invention have been described. However, it should be understood that various modifications can be made without departing from the spirit and scope of the present invention. Therefore, the descriptions in the experimental section and examples are intended to illustrate rather than limit the scope of the invention described in the claims.

[0180] 5.3 Protein

[0181] VZV glycoprotein includes glycoprotein E (gE), glycoprotein B (gB), glycoprotein H (gH) and glycoprotein L (gL), wherein gE is the most abundant VSV glycoprotein expressed by cells infected with VZV. VZV gE is generally 623 amino acid residues in length and comprises a signal peptide (usually amino acid residues 1-30 of full-length gE; For example, the signal peptide of GenBank accession number AAG32558.1 is composed of amino acid residues 1-30 of GenBank accession number AAG32558.1) and a transmembrane domain. Mature gE lacks a signal peptide. VZV gE forms a heterodimer with gI, and the heterodimer gE / gI is necessary for the intercellular spread of the virus. In addition, the VZVgE / gI heterodimer interacts with the Fc region of IgG. VZV gE is bound 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 Nos. 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 and Uniprot Nos. Q9J3M8 and P09259. In some embodiments, the VZV gE is gE of the Dumas strain. In some embodiments, the VZV gE is gE of the KPZ13-287 strain. In some embodiments, the VZV gE is gE of the VZVi / Munich.GER / 30.07 / Z[3] strain. In some embodiments, the VZV gE is gE of the NSYY3 strain. In some embodiments, the VZV gE is gE of the 1002 / 2008 strain. In some embodiments, the VZV gE is gE of the NSYY3 strain. In some embodiments, the VZV gE is gE of the Oka strain.

[0182] In some embodiments, provided herein are fragments of mature VZV gE, wherein the fragment comprises a truncation of at least one and at most 50, 49, 48, 47, 46, 45, 44, 43, or 42 amino acid residues from the C-terminus of mature gE. In some embodiments, the truncation is a truncation 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 truncation is a truncation 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 truncation is a truncation 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 truncation is a truncation of 14 or 37 amino acid residues from the C-terminus of mature gE. In some embodiments, the truncation is a truncation of 37 amino acid residues from the C-terminus of mature gE. In some embodiments, the truncation is a truncation 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, truncation is the truncation of up to 30 or 29 amino acid residues from the C-terminus of mature gE. In some embodiments, truncation is the truncation of up to 28 amino acid residues from the C-terminus of mature gE. In some embodiments, truncation is the truncation of up to 27 or 26 amino acid residues from the C-terminus of mature gE. In some embodiments, truncation is the truncation 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 1 amino acid residue from the C-terminus of mature gE. In some embodiments, mature gE comprises the amino acid sequence shown in SEQ ID NO:1.

[0183] In some embodiments, the fragment of mature gE further comprises one or more amino acid substitutions. In some embodiments, the fragment of mature gE further comprises one, two, three, four, five, or all of the amino acid substitutions selected from Y569A, Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residue positions 569, 582, 593, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55.

[0184] In some embodiments, the fragments of mature gE further comprise the amino acid residue substitution Y569A, wherein amino acid residue position number 569 is numbered according to the amino acid residue position of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise the amino acid residue substitution Y582G, wherein amino acid residue position 582 is numbered according to the amino acid residue position of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise the amino acid residue substitutions Y569A and Y582G, wherein amino acid residue positions 569 and 582 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise the amino acid residue substitutions Y569A, Y582G, and S593A, wherein amino acid residue positions 569, 582, and 593 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, Y582G, S595A, wherein amino acid residue positions 569, 582, and 595 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, Y582G, and T596A, wherein amino acid residue positions 569, 582, and 596 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, Y582G, and T598A, wherein amino acid residue positions 569, 582, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, Y582G, S593A, and S595A, wherein amino acid residue positions 569, 582, 593, and 595 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, wherein amino acid residue positions 569, 582, 593, 595, and 596 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residue positions 569, 582, 593, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55.

[0185] In some embodiments, the fragments of mature gE further comprise the amino acid residue substitution S593A, wherein amino acid residue position 593 is numbered according to the amino acid residue position of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise the amino acid residue substitution S595A, wherein amino acid residue position 595 is numbered according to the amino acid residue position of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise the amino acid residue substitution T596A, wherein amino acid residue position 596 is numbered according to the amino acid residue position of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise the amino acid residue substitution T598A, wherein amino acid residue position 598 is numbered according to the amino acid residue position of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise the amino acid residue substitutions S593A and S595A, wherein amino acid residue positions 593 and 595 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions S593A and T596A, wherein amino acid residue positions 593 and 596 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions S593A and T598A, wherein amino acid residue positions 593 and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions S595A and T596A, wherein amino acid residue positions 595 and 596 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions S595A and T598A, wherein amino acid residue positions 595 and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions T596A and T598A, wherein amino acid residue positions 596 and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions S593A, S595A, and T596A, wherein amino acid residue positions 593, 595, and 596 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions S593A, S595A, and T598A, wherein amino acid residue positions 593, 595, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions S595A, T596A, and T598A, wherein amino acid residue positions 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE.In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions S593A, S595A, T596A, T598A, wherein amino acid residue positions 593, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residue positions 582, 593, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55.

[0186] In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A and S593A, wherein amino acid residue positions 569 and 593 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A and S595A, wherein amino acid residue positions 569 and 595 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A and T596A, wherein amino acid residue positions 569 and 596 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A and T598A, wherein amino acid residue positions 569 and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A, S593A, and S595A, wherein amino acid residue positions 569, 593, and 595 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A, S593A, and T596A, wherein amino acid residue positions 569, 593, and 596 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A, S593A, and T598A, wherein amino acid residue positions 569, 593, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, S595A, and T596A, wherein amino acid residue positions 569, 595, and 596 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, S595A, and T598A, wherein amino acid residue positions 569, 595, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, T596A, and T598A, wherein amino acid residue positions 569, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragment of mature gE further comprises amino acid residue substitutions Y569A, S593A, S595A, and T596A, wherein amino acid residue positions 569, 593, 595, and 596 are numbered according to the amino acid residue positions of full-length VZV gE.In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A, S593A, S595A, and T598A, wherein amino acid residue positions 569, 593, 595, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A, S595A, T596A, T598A, wherein amino acid residue positions 569, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the fragments of mature gE further comprise amino acid residue substitutions Y569A, S593A, S595A, T596A, T598A, wherein amino acid residue positions 569, 593, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO:55.

[0187] 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 set forth 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 set forth 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 set forth 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 set forth 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 shown 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 shown in SEQ ID NO: 23. In some embodiments, the fragment of mature gE is a fragment described herein (e.g., a fragment of mature gE described in the Examples in this section or Section 6). In some embodiments, the fragment of mature gE comprises the amino acid sequence of SEQ ID NO: 3, 6, 8, 10, or 12. In specific embodiments, the fragment comprises the amino acid sequence of SEQ ID NO: 6. In some embodiments, the fragment of mature gE 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 fragment of mature gE 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 SEQ ID NO: 3, 6, 8, 10, or 12. In some embodiments, the fragment of mature gE 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 SEQ ID NO: 3, 6, 8, 10, or 12. In some embodiments, the fragment of mature gE 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 SEQ ID NO: 3, 6, 8, 10, or 12.

[0188] In some embodiments, the present invention provides a fusion protein comprising a fragment of mature gE of VZV and a human IgE signal peptide, wherein the fragment comprises a truncation 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 present invention provides a fusion protein comprising a fragment of mature gE of VZV and a human IgE signal peptide, wherein the fragment comprises a truncation of 37 amino acid residues from the C-terminus of mature gE and amino acid residue substitutions Y569A and Y582G, wherein amino acid residue positions 569 and 582 are numbered according to the amino acid residue positions of the 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 shown 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 shown 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 shown in SEQ ID NO:23.

[0189] In some embodiments, provided herein is a fusion protein comprising a fragment of mature gE of VZV and a human IgE signal peptide, wherein the fragment comprises a truncation of the C-terminal 37 amino acid residues of mature gE and the amino acid residue substitutions Y569A and Y582G, wherein amino acid residues 569 and 582 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, mature gE comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the fragment comprises the amino acid sequence set forth in SEQ ID NO: 6. In some embodiments, the human IgE signal peptide comprises the amino acid sequence set forth 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 set forth 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 shown in SEQ ID NO: 23. In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 59. In some embodiments, the fusion protein 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 of SEQ ID NO: 59. In some embodiments, the fusion protein 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 shown in SEQ ID NO: 59.

[0190] In some embodiments, provided herein is a fusion protein comprising a mutant of mature gE of VZV and a human IgE signal peptide, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residues 569, 582, 593, 595, 596, and 598 are numbered according to the amino acid residue positions 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 of SEQ ID NO: 55. In some embodiments, the mutant comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the human IgE signal peptide comprises the amino acid sequence set forth 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 set forth 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 shown in SEQ ID NO:23.

[0191] In some embodiments, provided herein is a fusion protein comprising a mutant of mature gE of VZV and a human IgE signal peptide, wherein the mutant comprises amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residues 582, 593, 595, 596, and 598 are numbered according to the amino acid residue positions of full-length VZV gE. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the mutant comprises the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the human IgE signal peptide comprises the amino acid sequence set forth 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 set forth 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 shown in SEQ ID NO:23.

[0192] In some embodiments, provided herein is a fusion protein comprising a fragment of mature gE of VZV and a human IgE signal peptide, wherein the fragment comprises a truncation of the C-terminal 50 amino acid residues of mature gE and the amino acid residue substitution Y569A, wherein amino acid residue 569 is numbered according to the amino acid residue position of full-length VZV gE. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO:55. In some embodiments, mature gE comprises the amino acid sequence set forth in SEQ ID NO:1. In some embodiments, the fragment comprises the amino acid sequence set forth in SEQ ID NO:3. In some embodiments, the human IgE signal peptide comprises the amino acid sequence set forth 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 set forth 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 shown in SEQ ID NO:23.

[0193] In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A and Y582G relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, and S593A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, and S595A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, and T596A relative to the full-length VSV gE.

[0194] In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, and T598A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, S593A, and S595A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, S593A, and T596A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, S593A, and T598A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, S595A, and T596A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, S595A, and T598A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, T596A, and T598A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of the full-length gE of VZV, wherein the mutant comprises amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A relative to the full-length VSV gE. In some embodiments, provided herein are mutants of full-length gE of VZV, wherein the mutants comprise amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A relative to full-length VSV gE. In some embodiments, full-length VZV gE comprises the amino acid sequence shown in SEQ ID NO: 55.

[0195] In some embodiments, provided herein is a protein comprising a mutant of mature gE of VZV, wherein the mutant comprises (i) a truncation of 37 amino acid residues from the C-terminus of mature gE, and (ii) amino acid residue substitutions Y569A and Y582G, wherein 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 set forth in SEQ ID NO: 1. In some embodiments, full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the mutant comprises the amino acid sequence set forth in SEQ ID NO: 6. In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO: 6. In some embodiments, the mutant 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 set forth in SEQ ID NO: 6. In some embodiments, the mutant 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 set forth in SEQ ID NO:6. In some embodiments, the mutant 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 set forth in SEQ ID NO:6. In some embodiments, the mutant 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 set forth in SEQ ID NO:6. In some embodiments, the protein further comprises a signal peptide of VZV gE. In some embodiments, the signal peptide of VZV gE comprises the amino acid sequence set forth in SEQ ID NO:18. In some embodiments, the signal peptide of VZV gE 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 set forth in SEQ ID NO:18. In some embodiments, the protein further comprises a heterologous signal peptide, wherein the N-terminus of the mutant 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 comprises the amino acid sequence set forth 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 set forth in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO: 23.In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO:6, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO:23. In some embodiments, the protein comprises the amino acid sequence set forth in SEQ ID NO:59. In some embodiments, the amino acid sequence of the protein consists of the amino acid sequence set forth in SEQ ID NO:59. In some embodiments, the heterologous signal peptide of the protein is the human tPA signal peptide. In some embodiments, the human tPA signal peptide comprises the amino acid sequence set forth 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 set forth in SEQ ID NO:28.

[0196] In some embodiments, provided herein is a protein comprising a mutant of mature gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A and Y582G, wherein amino acid residue positions 569 and 582 are numbered as amino acid residue positions of full-length VSV 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 set forth in SEQ ID NO: 55. In some embodiments, the mutant comprises the amino acid sequence set forth in SEQ ID NO: 8. In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO: 8. In some embodiments, the mutant 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 set forth in SEQ ID NO: 8. In some embodiments, the mutant 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 set forth in SEQ ID NO: 8. In some embodiments, the mutant 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 set forth in SEQ ID NO:8. In some embodiments, the mutant 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 set forth in SEQ ID NO:8. 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 set forth 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 set forth in SEQ ID NO:18. In some embodiments, the protein further comprises a heterologous signal peptide, wherein the N-terminus of the mutant 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 comprises the amino acid sequence set forth 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 set forth in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO: 23.In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO:8, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO:23. In some embodiments, the heterologous signal peptide of the protein is the human tPA signal peptide. In some embodiments, the human tPA signal peptide comprises the amino acid sequence set forth 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 set forth in SEQ ID NO:28.

[0197] In some embodiments, provided herein is a protein comprising a mutant of mature gE of VZV, wherein the mutant comprises amino acid residue substitutions Y569A, Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residue positions 569, 582, 593, 595, 596, and 598 are numbered as amino acid residue positions of full-length VSV gE. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the mutant comprises the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the mutant 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 set forth in SEQ ID NO: 10. In some embodiments, the mutant 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 set forth in SEQ ID NO: 10. In some embodiments, the mutant 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 set forth in SEQ ID NO: 10. In some embodiments, the mutant 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 set forth in SEQ ID NO: 10. In some embodiments, the protein further comprises a signal peptide of VZV gE. In some embodiments, the signal peptide of VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 18. In some embodiments, the signal peptide of VZV gE 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 set forth in SEQ ID NO: 18. In some embodiments, the protein further comprises a heterologous signal peptide, wherein the N-terminus of the mutant 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 comprises the amino acid sequence set forth 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 set forth in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO: 23.In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO: 10, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO: 23. In some embodiments, the heterologous signal peptide of the protein is the human tPA signal peptide. In some embodiments, the human tPA signal peptide comprises the amino acid sequence set forth 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 set forth in SEQ ID NO: 28.

[0198] In some embodiments, provided herein is a protein comprising a mutant of mature gE of VZV, wherein the mutant comprises amino acid residue substitutions Y582G, S593A, S595A, T596A, and T598A, wherein amino acid residue positions 582, 593, 595, 596, and 598 are numbered as amino acid residue positions of full-length VSV gE. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the mutant comprises the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the mutant 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 set forth in SEQ ID NO: 12. In some embodiments, the mutant 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 set forth in SEQ ID NO: 12. In some embodiments, the mutant 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 set forth in SEQ ID NO: 12. In some embodiments, the mutant 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 set forth in SEQ ID NO: 12. In some embodiments, the protein further comprises a signal peptide of VZV gE. In some embodiments, the signal peptide of VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 18. In some embodiments, the signal peptide of VZV gE 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 set forth in SEQ ID NO: 18. In some embodiments, the protein further comprises a heterologous signal peptide, wherein the N-terminus of the mutant 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 comprises the amino acid sequence set forth 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 set forth in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO: 23.In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO: 12, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO: 23. In some embodiments, the heterologous signal peptide of the protein is the human tPA signal peptide. In some embodiments, the human tPA signal peptide comprises the amino acid sequence set forth 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 set forth in SEQ ID NO: 28.

[0199] In some embodiments, the present invention provides a protein comprising a mutant of mature gE of VZV, wherein the mutant comprises (i) a truncation of the C-terminal 50 amino acid residues of the mature gE protein, and (ii) an amino acid residue substitution Y569A, wherein amino acid residue position 569 is the amino acid residue position of the full-length VSV gE. In some embodiments, the mature gE comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the full-length VZV gE comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the mutant comprises the amino acid sequence set forth in SEQ ID NO: 3. In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO: 3. In some embodiments, the mutant 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 set forth in SEQ ID NO: 3. In some embodiments, the mutant 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 set forth in SEQ ID NO: 3. In some embodiments, the mutant 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 set forth in SEQ ID NO:3. In some embodiments, the mutant 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 set forth in SEQ ID NO:3. In some embodiments, the protein further comprises a VZV gE signal peptide comprising the amino acid sequence set forth 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 set forth in SEQ ID NO:18. In some embodiments, the protein further comprises a heterologous signal peptide, wherein the N-terminus of the mutant 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 comprises the amino acid sequence set forth 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 shown in SEQ ID NO: 23. In some embodiments, the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence shown in SEQ ID NO: 23.In some embodiments, the amino acid sequence of the mutant consists of the amino acid sequence set forth in SEQ ID NO:3, and the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence set forth in SEQ ID NO:23. In some embodiments, the heterologous signal peptide of the protein is the human tPA signal peptide. In some embodiments, the human tPA signal peptide comprises the amino acid sequence set forth 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 set forth in SEQ ID NO:28.

[0200] In some embodiments, provided herein is a protein comprising the amino acid sequence of a VZV gE mutant protein disclosed in Table 1. In some embodiments, provided herein is a protein comprising the amino acid sequence of a VZV gE mutant protein-1 disclosed in Table 1. In some embodiments, provided herein is a protein comprising the amino acid sequence of a VZV gE mutant protein-2 disclosed in Table 1. In some embodiments, provided herein is a protein comprising the amino acid sequence of a VZV gE mutant protein-3 disclosed in Table 1. In some embodiments, provided herein is a protein comprising the amino acid sequence of a VZV gE mutant protein-4 disclosed in Table 1. In some embodiments, provided herein is a protein comprising the amino acid sequence of a VZV gE mutant protein-5 disclosed in Table 1.

[0201] In some embodiments, provided herein are mutants of the VZV gE protein described below in Section 6. In some embodiments, provided herein are mutants of the VZV gE other than controls described below in Section 6. In some embodiments, provided herein are mutants of the VZV gE other than controls described below in Section 6, encoded by nucleic acids.

[0202] In some embodiments, the fragments described herein, the mutants described herein, or the 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, for example, X-ray crystallography, nuclear magnetic resonance), or binding to an antibody specific for a conformational epitope of wild-type VZV gE. In some embodiments, the proteins comprising the mutants of mature gE described herein have a structure similar to wild-type VZV gE, as assessed by techniques known to those skilled in the art (such as, for example, X-ray crystallography, nuclear magnetic resonance), or binding to an antibody specific for a conformational epitope of wild-type VZV gE.

[0203] In some embodiments, the fragments, mutants, or fusion proteins described herein retain at least one activity or function of mature VZV gE. In some embodiments, the proteins comprising the mutants of mature gE described herein 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 another example, in some embodiments, the ability to bind to Fc receptors is retained. In another example, in some embodiments, the ability to bind to insulin degrading enzyme (IDE) is retained. In another example, in some embodiments, the ability to be phosphorylated by the viral kinase encoded by ORF47 is retained.

[0204] 5.4 Therapeutic Nucleic Acids

[0205] In one aspect, provided herein are therapeutic nucleic acid molecules for the management, prevention, and treatment of VZV infection. In some embodiments, the therapeutic nucleic acid encodes a peptide or polypeptide that, when administered to a subject in need thereof, is expressed by cells in the subject to produce 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 specific embodiments, the therapeutic nucleic acid molecule is an mRNA molecule.

[0206] In some embodiments, the therapeutic nucleic acid molecule is formulated in a vaccine composition. In some embodiments, the vaccine composition is a genetic vaccine as described herein. In some embodiments, the vaccine composition comprises an mRNA molecule as described herein.

[0207] In some embodiments, the mRNA molecules of the present disclosure encode target peptides or polypeptides, including any naturally occurring or non-naturally occurring or otherwise modified polypeptides. The peptides or polypeptides encoded by the mRNA can be of any size and can have any secondary structure or activity. In some embodiments, the polypeptides encoded by the mRNA payload can have a therapeutic effect when expressed in a cell.

[0208] In some embodiments, the mRNA molecules of the present disclosure include at least one coding region (e.g., open reading frame (ORF)) encoding a target peptide or polypeptide. In some embodiments, the nucleic acid molecule further includes at least one untranslated region (UTR). In specific embodiments, the untranslated region (UTR) is located upstream (5' end) of the coding region and is referred to herein as a 5'-UTR. In specific embodiments, the untranslated region (UTR) is located downstream (3' end) of the coding region and is referred to herein as a 3'-UTR. In specific embodiments, the nucleic acid molecule includes both a 5'-UTR and a 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 a 5'-UTR). In some embodiments, the nucleic acid molecule includes a poly-A region (e.g., in a 3'-UTR). In some embodiments, the nucleic acid molecule includes a polyadenylation signal (e.g., in a 3'-UTR). In some embodiments, the nucleic acid molecule includes a stabilizing region (e.g., in a 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 comprises a stem-loop sequence (e.g., in a 5'-UTR and / or a 3'-UTR). In some embodiments, the nucleic acid molecule comprises one or more intronic regions that can be excised during splicing. In specific embodiments, the nucleic acid molecule comprises one or more regions selected from a 5'-UTR and a coding region. In specific embodiments, the nucleic acid molecule comprises one or more regions selected from a coding region and a 3'-UTR. In specific embodiments, the nucleic acid molecule comprises one or more regions selected from a 5'-UTR, a coding region, and a 3'-UTR.

[0209] 5.4.1 Coding Region

[0210] In some embodiments, 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 ORF encoding a peptide or protein. In embodiments where the coding region includes more than one ORF, the encoded peptides and / or proteins may be identical or different from each other. In some embodiments, the multiple ORFs in the coding region are separated by non-coding sequences. In specific embodiments, the non-coding sequence separating two ORFs includes an internal ribosome entry site (IRES).

[0211] Without being bound by theory, it is expected that an internal ribosome entry site (IRES) can serve as the sole ribosome binding site, or as one of multiple ribosome binding sites of an mRNA. An mRNA molecule containing more than one functional ribosome binding site can encode several peptides or proteins that are independently translated by the ribosome (e.g., polycistronic mRNA). Thus, in some embodiments, the nucleic acid molecules (e.g., mRNA) of the present disclosure comprise one or more internal ribosome entry sites (IRES). Examples of IRES sequences that can be used in conjunction with the present disclosure include, but are not limited to, those from picornaviruses (e.g., FMDV), pestiviruses (CFFV), polioviruses (PV), encephalomyocarditis virus (ECMV), foot-and-mouth disease virus (FMDV), hepatitis C virus (HCV), classical swine fever virus (CSFV), murine leukemia virus (MLV), simian immunodeficiency virus (SIV), or cricket paralysis virus (CrPV).

[0212] In various embodiments, the nucleic acid molecules of the present disclosure encode at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more 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 present disclosure encode dipeptides (e.g., carnosine 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 molecules encode peptides or polypeptides having at least about 150 amino acids. In some embodiments, the nucleic acid molecule encoding has a peptide or polypeptide of at least about 300 amino acids. In some embodiments, the nucleic acid molecule encoding has a peptide or polypeptide of at least about 500 amino acids. In some embodiments, the nucleic acid molecule encoding has a peptide or polypeptide of at least about 1000 amino acids.

[0213] In some embodiments, the length of the nucleic acid molecules of the present disclosure is at least about 30 nucleotides (nt). In some embodiments, the length of the nucleic acid molecules is at least about 35nt. In some embodiments, the length of the nucleic acid molecules is at least about 40nt. In some embodiments, the length of the nucleic acid molecules is at least about 45nt. In some embodiments, the length of the nucleic acid molecules is at least about 50nt. In some embodiments, the length of the nucleic acid molecules is at least about 55nt. In some embodiments, the length of the nucleic acid molecules is at least about 60nt. In some embodiments, the length of the nucleic acid molecules is at least about 65nt. In some embodiments, the length of the nucleic acid molecules is at least about 70nt. In some embodiments, the length of the nucleic acid molecules is at least about 75nt. In some embodiments, the length of the nucleic acid molecules is at least about 80nt. In some embodiments, the length of the nucleic acid molecules is at least about 85nt. In some embodiments, the length of the nucleic acid molecules is at least about 90nt. In some embodiments, the length of the nucleic acid molecules is at least about 95nt. In some embodiments, the length of the nucleic acid molecules is at least about 100nt. In some embodiments, the length of the nucleic acid molecules is at least about 120nt. In some embodiments, the length of the nucleic acid molecule is at least about 140 nt. In some embodiments, the length of the nucleic acid molecule is at least about 160 nt. In some embodiments, the length of the nucleic acid molecule is at least about 180 nt. In some embodiments, the length of the nucleic acid molecule is at least about 200 nt. In some embodiments, the length of the nucleic acid molecule is at least about 250 nt. In some embodiments, the length of the nucleic acid molecule is at least about 300 nt. In some embodiments, the length of the nucleic acid molecule is at least about 400 nt. In some embodiments, the length of the nucleic acid molecule is at least about 500 nt. In some embodiments, the length of the nucleic acid molecule is at least about 600 nt. In some embodiments, the length of the nucleic acid molecule is at least about 700 nt. In some embodiments, the length of the nucleic acid molecule is at least about 800 nt. In some embodiments, the length of the nucleic acid molecule is at least about 900 nt. In some embodiments, the length of the nucleic acid molecule is at least about 1000 nt. In some embodiments, the length of the nucleic acid molecule is at least about 1100 nt. In some embodiments, the length of the nucleic acid molecule is at least about 1200 nt. In some embodiments, the length of the nucleic acid molecule is at least about 1300 nt. In some embodiments, the length of the nucleic acid molecule is at least about 1400nt. In some embodiments, the length of the nucleic acid molecule is at least about 1500nt. In some embodiments, the length of the nucleic acid molecule is at least about 1600nt. In some embodiments, the length of the nucleic acid molecule is at least about 1700nt. In some embodiments, the length of the nucleic acid molecule is at least about 1800nt. In some embodiments, the length of the nucleic acid molecule is at least about 1900nt.In some embodiments, the length of the nucleic acid molecule is at least about 2000nt. In some embodiments, the length of the nucleic acid molecule is at least about 2500nt. In some embodiments, the length of the nucleic acid molecule is at least about 3000nt. In some embodiments, the length of the nucleic acid molecule is at least about 3500nt. In some embodiments, the length of the nucleic acid molecule is at least about 4000nt. In some embodiments, the length of the nucleic acid molecule is at least about 4500nt. In some embodiments, the length of the nucleic acid molecule is at least about 5000nt.

[0214] In a specific embodiment, the therapeutic nucleic acids of the present disclosure are formulated as vaccine compositions as described herein (e.g., gene vaccines). In some embodiments, the therapeutic nucleic acid encoding peptides or proteins that can induce immunity against one or more target disorders or diseases. In some embodiments, the target disorder is associated with or caused by a pathogen infection, such as VZV. In some embodiments, the therapeutic nucleic acid sequence (e.g., mRNA) encodes a pathogen-characteristic pathogenic protein or an immunogenic fragment (e.g., epitope) or derivative thereof. After being administered to a vaccinated subject, the vaccine allows the expression of the encoded pathogenic protein (or its immunogenic fragment or derivative), thereby inducing immunity against the pathogen in the subject.

[0215] In specific embodiments, provided herein are therapeutic compositions (eg, vaccine compositions) for the management, prevention, and treatment of diseases or conditions caused by VZV or infection with VZV.

[0216] Without being bound by theory, consider VZV, i.e. varicella-zoster virus, also known as human herpesvirus type 3, a double-stranded DNA virus belonging to the alpha herpesvirus class. VZV has only one serotype. VZV has a genome comprising 71 genes and encoding 67 proteins, including 6 glycoproteins, now designated gE, gB, gH, gI, gC, and gL. Glycoproteins gE, gB, and gH are highly abundant in infected cells and are also present in the envelope of virions. Antibodies induced by the three major glycoproteins can neutralize the virus. Specific humoral and cellular immunity, as well as cytokines such as interferon, play a major role in limiting the spread and recovery of VZV, with specific cellular immunity being particularly important.

[0217] Thus, in some embodiments, provided herein are therapeutic nucleic acids encoding viral peptides or proteins derived from VZV. In some embodiments, the nucleic acid encodes a viral peptide or protein derived from VZV, wherein the viral peptide or protein is one or more selected from the following: (a) gE protein; (b) gB protein; (c) gH protein; (d) gI protein; (e) gC protein; (f) gL protein; (g) an immunogenic fragment of any one of (a) to (f); and (h) a functional derivative of any one of (a) to (g).

[0218] Thus, in some embodiments, the therapeutic nucleic acids of the present disclosure encode VZV gE protein, or an immunogenic fragment of a gE protein, or a functional derivative of a gE protein or an immunogenic fragment thereof. Table 1 shows, among other things, exemplary VZV native antigen sequences.

[0219] Table 1. Exemplary sequences of VZV antigens.

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233] Note: Sequences in italics (in case of full-length protein) are signal peptides.

[0234] In specific embodiments, the therapeutic nucleic acids of the present disclosure encode the mature gE protein of VZV, wherein the mature gE protein has the amino acid sequence of SEQ ID NO: 1. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode the mature gE protein of VZV, wherein the therapeutic nucleic acid comprises the DNA coding sequence of SEQ ID NO: 2. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode the mature gE protein of VZV, wherein the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 2. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode the full-length gE protein of VZV, wherein the full-length gE protein has the amino acid sequence of SEQ ID NO: 55. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode the full-length gE protein of VZV, wherein the therapeutic nucleic acid comprises the DNA coding sequence of SEQ ID NO: 56. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode the full-length gE protein of VZV, wherein the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 56. In some embodiments, the RNA sequence is in vitro transcribed. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0235] In certain embodiments, the therapeutic nucleic acids of the present disclosure encode immunogenic fragments of the gE protein of VZV. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode functional derivatives of the gE protein of VZV. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode immunogenic fragments of the gE protein of VZV, wherein the immunogenic fragment of the gE protein comprises a truncation of at least 1 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 immunogenic fragments of the gE protein of VZV, wherein the immunogenic fragments of the gE protein comprise truncations 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 1 amino acid residue from the C-terminus compared to the mature gE protein. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode immunogenic fragments of the gE protein of VZV, wherein the immunogenic fragments of the gE protein comprise a truncation 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. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode immunogenic fragments of the gE protein of VZV, wherein the immunogenic fragments of the gE protein comprise a truncation 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 the mature gE protein. In certain embodiments, the therapeutic nucleic acid of the present disclosure encodes an immunogenic fragment of the gE protein of VZV, wherein the immunogenic fragment of the gE protein comprises a truncation of 14 or 37 amino acid residues from the C-terminus compared to the mature gE protein. In some embodiments, the RNA sequence is in vitro transcribed. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0236] In a specific embodiment, the therapeutic nucleic acid of the present invention encodes a mutant of the gE protein of VZV. In a specific embodiment, the therapeutic nucleic acid of the present invention encodes a mutant of the gE protein of VZV, wherein the mutant comprises the substitution Y569A. In a specific embodiment, the therapeutic nucleic acid of the present invention encodes a mutant of the gE protein of VZV, wherein the mutant comprises the substitution Y582G. In a specific embodiment, the therapeutic nucleic acid of the present invention encodes a mutant of the gE protein of VZV, wherein the mutant comprises the substitution Y569A and Y582G. In a specific embodiment, the therapeutic nucleic acid of the present invention encodes a mutant of the gE protein of VZV, wherein the mutant comprises the substitution S593A. In a specific embodiment, the therapeutic nucleic acid of the present invention encodes a mutant of the gE protein of VZV, wherein the mutant comprises the substitution S595A. In a specific embodiment, the therapeutic nucleic acid of the present invention encodes a mutant of the gE protein of VZV, wherein the mutant comprises the substitution T596A. In a specific embodiment, the therapeutic nucleic acid of the present disclosure encodes a mutant of the gE protein of VZV, wherein the mutant comprises the substitution T598A. In a specific embodiment, the therapeutic nucleic acid of the present disclosure encodes a mutant of the gE protein of VZV, wherein the mutant comprises the substitutions S593A, S595A, T596A, and T598A. In a specific embodiment, the therapeutic nucleic acid of the present disclosure encodes a mutant of the gE protein of VZV, wherein the mutant comprises the 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 in vitro transcribed. In a specific embodiment, the nucleic acid molecule is an mRNA molecule.

[0237] In a specific embodiment, the therapeutic nucleic acid encoding VZV of the present invention is a mutant fragment of the gE protein. In a specific embodiment, the therapeutic nucleic acid encoding VZV of the present invention is a mutant fragment of the gE protein, wherein compared with the mature gE protein, the mutant fragment comprises a truncation of 49, 48, 47, 46, 45, 44, 43 or 42 amino acid residues from the C-terminus. In such embodiments, the mutant fragment optionally comprises the substitution Y569A. In a specific embodiment, the therapeutic nucleic acid encoding VZV of the present invention is a mutant fragment of the gE protein, wherein compared with the mature gE protein, the mutant fragment comprises the truncation of 41, 40, 39, 38, 37, 36, 35, 34, 33, 32 or 31 amino acid residues from the C-terminus. In such embodiments, the mutant fragment optionally comprises the substitution Y582G, with or without the substitution Y569A. In such embodiments, the mutant fragment comprises the substitutions Y569A and Y582G. In a specific embodiment, the therapeutic nucleic acid encoding VZV of the present invention is a mutant fragment of the gE protein, wherein the mutant fragment comprises a truncation 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 a substitution of S593A, with or without substitutions of Y569A and / or Y582G. In such embodiments, the mutant fragment comprises substitutions of Y569A, Y582G and S593A. In a specific embodiment, the therapeutic nucleic acid encoding VZV of the present invention is a mutant fragment of the gE protein, wherein the mutant fragment comprises a truncation of 28 amino acid residues from the C-terminus compared to the mature gE protein. In such embodiments, the mutant fragment optionally comprises a substitution of S595A, with or without substitutions of Y569A and / or Y582G and / or S593A. In such embodiments, the mutant fragment comprises substitutions of Y569A, Y582G, S593A and S595A. In certain embodiments, the therapeutic nucleic acid of the present disclosure encodes a mutant fragment of the gE protein of VZV, wherein the mutant fragment comprises a truncation 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 the substitution T596A, with or without the substitutions Y569A and / or Y582G and / or S593A and / or S595A. In such embodiments, the mutant fragment comprises the substitutions Y569A, Y582G, S593A, S595A, and T596A. In certain embodiments, the therapeutic nucleic acid of the present disclosure encodes a mutant fragment of the gE protein of VZV, wherein the mutant fragment comprises a truncation 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 from the C-terminus compared to the mature gE protein.In such embodiments, the mutant fragment optionally comprises the substitution T598A, with or without the substitutions Y569A and / or Y582G and / or S593A and / or S595A and / or T596A. In such embodiments, the mutant fragment comprises the substitutions Y569A, Y582G, S593A, S595A, T596A and T598A. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the mutant fragment has the amino acid sequence of SEQ ID NO: 3. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises the DNA coding sequence of SEQ ID NO: 4 or 5. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 4 or 5. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the mutant fragment has the amino acid sequence of SEQ ID NO: 6. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises the DNA coding sequence of SEQ ID NO: 7. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 7. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the mutant fragment has the amino acid sequence of SEQ ID NO: 8. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises the DNA coding sequence of SEQ ID NO: 9. In specific embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 9. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the mutant fragment has the amino acid sequence of SEQ ID NO: 10. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises the DNA coding sequence of SEQ ID NO: 11. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 11. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the mutant fragment has the amino acid sequence of SEQ ID NO: 12.In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises the DNA coding sequence of SEQ ID NO: 13. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a mutant fragment of the gE protein of VZV, wherein the therapeutic nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence of SEQ ID NO: 13. In some embodiments, the RNA sequence is transcribed in vitro. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0238] Without being bound by theory, it is contemplated that 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 trimerizing peptide such that the fusion protein is capable of forming a trimeric complex comprising three copies of the gE protein or fragment thereof. In some embodiments, the gE protein or fragment thereof is fused to the trimerizing peptide via a peptide linker.

[0239] Table 2 shows the sequences of exemplary trimerization peptides and linker peptides, as well as fusion proteins that can be used in conjunction with the present disclosure.

[0240] Table 2. Exemplary sequences of linker peptides and trimerization peptides.

[0241]

[0242] In some embodiments, the therapeutic nucleic acid encodes a fusion protein comprising the VZV gE protein, or a functional derivative thereof, fused to a trimerizing peptide. In some embodiments, the fusion between the gE protein and the trimerizing peptide is via a peptide linker. In specific embodiments, the peptide linker comprises the amino acid sequence of SEQ ID NO: 14. In some embodiments, the trimerizing peptide comprises the amino acid sequence of SEQ ID NO: 16.

[0243] In certain embodiments, the therapeutic nucleic acid encodes a fusion protein comprising the VZV gE protein fused to a trimerizing peptide, wherein the nucleic acid comprises a DNA coding sequence. In certain embodiments, the therapeutic nucleic acid encodes a fusion protein comprising the VZV gE protein fused to a trimerizing peptide, wherein the nucleic acid comprises an RNA sequence transcribed from the DNA coding sequence. In some embodiments, the RNA sequence is in vitro transcribed. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0244] Without being bound by theory, it is expected that a fusion protein comprising a viral peptide or polypeptide fused to an immunoglobulin Fc region can enhance the immunogenicity of the viral peptide or polypeptide. Therefore, in some embodiments, the therapeutic nucleic acid molecules of the present disclosure encode a fusion protein comprising a viral peptide or protein derived from VZV fused to the Fc region of an immunoglobulin. In specific embodiments, the viral peptide or protein is one or more selected from the following: (a) gE protein; (b) gB protein; (c) gH protein; (d) gI protein; (e) gC protein; (f) gL protein; (g) an immunogenic fragment of any one of (a) to (f); and a functional derivative of any one of (h) (a) to (g). In specific embodiments, the immunoglobulin is a human immunoglobulin (Ig). In specific embodiments, the immunoglobulin is human IgG, IgA, IgD, IgE, or IgM. In specific embodiments, the immunoglobulin is human IgG1, IgG2, IgG3, or IgG4. In some embodiments, the immunoglobulin Fc is fused to the N-terminus of the viral peptide or polypeptide. In other embodiments, the immunoglobulin Fc is fused to the C-terminus of the viral peptide or polypeptide.

[0245] Without being bound by theory, it is expected that the signal peptide can mediate the transport of the polypeptide fused thereto to a specific location of the 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 specific embodiments, the viral peptide or protein is one or more selected from the following: (a) gE protein; (b) gB protein; (c) gH protein; (d) gI protein; (e) gC protein; (f) gL protein; (g) an immunogenic fragment of any one of (a) to (f); and (h) a functional derivative of any one of (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 conjunction with the present disclosure, as well as exemplary VZV antigen sequences comprising the signal peptides.

[0246] Table 3: Exemplary sequences of signal peptides.

[0247]

[0248]

[0249] In certain embodiments, the signal peptide is encoded by the gene of VZV from which the viral peptide or polypeptide is derived. In certain embodiments, the signal peptide encoded by the gene of VZV is fused to a viral peptide or polypeptide encoded by a different gene of VZV. In other embodiments, the signal peptide encoded by the gene of VZV is fused to a viral peptide or polypeptide encoded by the same gene of VZV. For example, in some embodiments, a signal peptide having the amino acid sequence of 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 the following: (a) E protein; (b) B protein; (c) H protein; (d) I protein; (e) C protein; (f) L protein; (g) an immunogenic fragment of any one of (a) to (f); and (h) a functional derivative of any one of (a) to (g).

[0250] In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a VZV gE protein or fragment without a native signal peptide. In certain embodiments, the encoded gE protein or fragment comprises a signal peptide having an amino acid sequence of SEQ ID NO: 23 or 27. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a VZV gE protein or fragment with a signal peptide, and wherein the therapeutic nucleic acid comprises a DNA coding sequence of SEQ ID NO: 24, 25, 26, or 28. In certain embodiments, the therapeutic nucleic acids of the present disclosure encode a VZV gE protein or fragment with a signal peptide, and wherein the therapeutic nucleic acid comprises an RNA sequence transcribed from a DNA coding sequence of SEQ ID NO: 24, 25, 26, or 28. In some embodiments, the RNA sequence is in vitro transcribed. In certain embodiments, the nucleic acid molecule is an mRNA molecule.

[0251] In other embodiments, the signal peptide is encoded by an exogenous gene sequence that is not present in the VZV from which the viral peptide or polypeptide is derived. In some embodiments, the heterologous signal peptide replaces the homologous signal peptide in the fusion protein encoded by the nucleic acid molecule of the present 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 with an amino acid sequence of MDWTWILFLVAAATRVHS (SEQ ID NO: 23) 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 the following: (a) E protein; (b) B protein; (c) H protein; (d) I protein; (e) C protein; (f) L protein; (g) an immunogenic fragment of any one of (a) to (f); and (h) a functional derivative of any one of (a) to (g).

[0252] In some embodiments, provided herein are nucleic acids encoding fragments of the mature gE described in Section 5.3. In some embodiments, provided herein are nucleic acids encoding the fusion proteins described in Section 5.3. In some embodiments, provided herein are nucleic acids encoding mutants of the full-length VZV gE described in Section 5.3. In some embodiments, provided herein are nucleic acids encoding proteins described in Section 5.3. In some embodiments, provided herein are nucleic acids comprising the nucleotide sequence set forth in SEQ ID NO: 7, 9, 11, 13, 4, or 5. In some embodiments, provided herein are nucleic acids comprising 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 set forth in SEQ ID NO: 7, 9, 11, 13, 4, or 5. In some embodiments, the nucleic acid is non-naturally occurring. 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 an mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5' untranslated region (5'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3' untranslated region (3'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as, for example, as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as, for example, as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, eg, as described in Section 5.4.4.

[0253] In some embodiments, provided herein are nucleic acids encoding the proteins or fusion proteins described in Section 5.3, comprising a mutant or fragment of mature gE and a human IgE signal peptide, wherein the nucleotide sequence encoding the IgE signal peptide comprises the nucleotide sequence set forth in SEQ ID NO: 24, 25, or 26. In some embodiments, provided herein are nucleic acids encoding the proteins or fusion proteins described in Section 5.3, comprising a mutant 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 set forth in SEQ ID NO: 24, 25, or 26. In some embodiments, the nucleic acid is non-naturally occurring. 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 an mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5' untranslated region (5'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3' untranslated region (3'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, such as described in Section 5.4.4.

[0254] In some embodiments, provided herein are nucleic acids encoding the proteins described in Section 5.3, comprising a mutant of mature gE and a signal peptide, wherein the nucleotide sequence encoding the signal peptide comprises the nucleotide sequence set forth in SEQ ID NO: 19, 20, 21, or 22. In some embodiments, provided herein are nucleic acids encoding the proteins described in Section 5.3, comprising a mutant 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 set forth in SEQ ID NO: 19, 20, 21, or 22. In some embodiments, the nucleic acid is non-naturally occurring. 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-pseudouridine and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid also comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid also comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid also comprises, for example, a 5'-untranslated region (5'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid also comprises, for example, a 3'-untranslated region (3'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid also comprises, for example, a 5'-untranslated region (5'-UTR) as described in sections 5.4.3 and, for example, a 3'-untranslated region (3'-UTR) as described in sections 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence shown 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, such as described in Section 5.4.4.

[0255] In some embodiments, provided herein are nucleic acids encoding the protein or fusion protein described in Section 5.3, comprising a mutant 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 set forth in SEQ ID NO: 28. In some embodiments, provided herein are nucleic acids encoding the protein or fusion protein described in Section 5.3, comprising a mutant 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 set forth in SEQ ID NO: 28. In some embodiments, the nucleic acid is non-naturally occurring. 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 an mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-untranslated region (5'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3'-untranslated region (3'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as, for example, as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as, for example, as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, eg, as described in Section 5.4.4.

[0256] In some embodiments, provided herein are nucleic acids comprising the nucleotide sequence set forth in SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, provided herein are nucleic acids comprising the nucleotide sequence set forth in SEQ ID NO: 63. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising the nucleotide sequence set forth in SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising the nucleotide sequence set forth in SEQ ID NO: 63. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 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 is non-naturally occurring. 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 an mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-untranslated region (5'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3'-untranslated region (3'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, such as described in Section 5.4.4.

[0257] In some embodiments, provided herein are nucleic acids encoding the amino acid sequence shown in SEQ ID NO: 6. In some embodiments, the nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 7. In some embodiments, provided herein are nucleic acids encoding the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 9. In some embodiments, provided herein are nucleic acids encoding the amino acid sequence shown in SEQ ID NO: 10. In some embodiments, the nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 11. In some embodiments, provided herein are nucleic acids encoding the amino acid sequence shown in SEQ ID NO: 12. In some embodiments, the nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 13. In some embodiments, provided herein are nucleic acids encoding the amino acid sequence shown in SEQ ID NO: 3. In some embodiments, the nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 4 or 5. In some embodiments, the nucleic acid is non-naturally occurring. 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 an mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-untranslated region (5'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3'-untranslated region (3'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46.In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, such as described in Section 5.4.4.

[0258] In some embodiments, provided herein are nucleic acids encoding the amino acid sequence of SEQ ID NO: 59. In some embodiments, the nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid comprises the nucleotide sequence shown in SEQ ID NO: 63. In some embodiments, the nucleic acid consists of the nucleotide sequence shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid consists of the nucleotide sequence shown 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 shown 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 shown 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 92%, at least 93%, or at least 94% identical to the nucleotide sequence shown 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 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the nucleotide sequence shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid is non-naturally occurring. 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-pseudouridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure, such as, for example, as described in Section 5.4.2. In some embodiments, the nucleic acid further comprises a 5'-cap structure, such as, for example, as described in Section 5.4.2. In some embodiments, the nucleic acid further comprises a 5'-untranslated region (5'-UTR), such as, for example, as described in Section 5.4.3. In some embodiments, the nucleic acid further comprises a 3'-untranslated region (3'-UTR), such as, for example, as described in Section 5.4.3.In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, such as described in Section 5.4.4.

[0259] In some embodiments, provided herein are nucleic acids comprising the nucleotide sequence shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, provided herein are nucleic acids consisting of the nucleotide sequence shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 51, 60, 61, 62, 63, or 64. In some embodiments, the nucleic acid is non-naturally occurring. 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 an mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-untranslated region (5'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3'-untranslated region (3'-UTR) as described in sections 5.4.3.In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, such as described in Section 5.4.4.

[0260] In some embodiments, provided herein are nucleic acids comprising the nucleotide sequence shown in SEQ ID NO: 63. In some embodiments, provided herein are nucleic acids consisting of the nucleotide sequence shown in SEQ ID NO: 63. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 63. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 63. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 63. In some embodiments, provided herein is a nucleic acid consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 63. In some embodiments, the nucleic acid is non-naturally occurring. 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-pseudouridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises a 5'-cap structure, such as described in Section 5.4.2. In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3. In some embodiments, the nucleic acid further comprises a 3' untranslated region (3'-UTR), such as described in Section 5.4.3. In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3 and a 3' untranslated region (3'-UTR), such as described in Section 5.4.3.In some embodiments, the 5'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence shown 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, such as described in Section 5.4.4.

[0261] In some embodiments, provided herein are nucleic acids comprising the nucleotide sequence shown in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, provided herein are nucleic acids consisting of the nucleotide sequence shown in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising a nucleotide sequence that is at least 90%, at least 91%, at least 92%, or at least 93% identical to 49, 50, 52, 53, or 5460, 61, 62, 63, or 64. In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising 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 shown in SEQ ID NO: 49, 50, 52, 53, or 54. In some embodiments, the nucleic acid is non-naturally occurring. 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 an mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in sections 5.4.9). In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-cap structure as described in sections 5.4.2. In some embodiments, the nucleic acid further comprises, for example, a 5'-untranslated region (5'-UTR) as described in sections 5.4.3. In some embodiments, the nucleic acid further comprises, for example, a 3'-untranslated region (3'-UTR) as described in sections 5.4.3.In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, such as described in Section 5.4.4.

[0262] In some embodiments, provided herein are nucleic acids shown in Table 1 or mRNAs for nucleic acids shown in Table 1. In some embodiments, provided herein are nucleic acids shown in Table 1 other than SEQ ID NOs: 2 and 56, or mRNAs for nucleic acids shown in Table 1 other than SEQ ID NOs: 2 and 56. In some embodiments, provided herein are nucleic acids comprising the VZV gE mutant protein coding sequences shown in Table 1, or mRNAs for nucleic acids comprising the VZV gE mutant protein coding sequences shown in Table 1. In some embodiments, provided herein are nucleic acids consisting of the VZV gE mutant protein coding sequences shown in Table 1, or mRNAs for nucleic acids comprising the VZV gE mutant protein coding sequences shown in Table 1. In some embodiments, provided herein are nucleic acids comprising the VZV gE mutant protein-1 coding sequences shown in Table 1, or mRNAs for nucleic acids comprising the VZV gE mutant protein-1 coding sequences shown in Table 1. In some embodiments, provided herein are nucleic acids comprising the VZV gE mutant protein-2 coding sequences shown in Table 1, or mRNAs for nucleic acids comprising the VZV gE mutant protein-2 coding sequences shown in Table 1. In some embodiments, provided herein are nucleic acids comprising a VZV gE mutant protein-3 coding sequence as shown in Table 1, or mRNAs of nucleic acids comprising a VZV gE mutant protein-3 coding sequence as shown in Table 1. In some embodiments, provided herein are nucleic acids comprising a VZV gE mutant protein-4 coding sequence as shown in Table 1, or mRNAs of nucleic acids comprising a VZV gE mutant protein-4 coding sequence as shown in Table 1. In some embodiments, provided herein are nucleic acids comprising a VZV gE mutant protein-5 coding sequence as shown in Table 1, or mRNAs of nucleic acids comprising a VZV gE mutant protein-5 coding sequence as shown 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 an mRNA, and the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine, and 5-methylcytosine. In some embodiments, the nucleic acid comprises one or more modified internucleoside linkages (e.g., one or more modified internucleoside linkages described in Section 5.4.9). In some embodiments, the nucleic acid further comprises a 5'-cap structure, such as, for example, as described in Section 5.4.2. In some embodiments, the nucleic acid further comprises a 5'-cap structure, such as, for example, as described in Section 5.4.2. In some embodiments, the nucleic acid further comprises a 5'-untranslated region (5'-UTR), such as, for example, as described in Section 5.4.3. In some embodiments, the nucleic acid further comprises a 3'-untranslated region (3'-UTR), such as, for example, as described in Section 5.4.3.In some embodiments, the nucleic acid further comprises a 5' untranslated region (5'-UTR), such as described in Section 5.4.3, and a 3' untranslated region (3'-UTR), such as described in Section 5.4.3. In some embodiments, the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38. In some embodiments, the 3'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 39-46. In some embodiments, the nucleic acid further comprises a 5'-UTR and a 3'-UTR, wherein the 5'-UTR comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 29-38, and the 3'-UTR comprises the nucleotide sequence set forth 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, such as described in Section 5.4.4.

[0263] In specific embodiments, the encoding nucleotide sequence of a nucleic acid described herein (e.g., a naturally occurring nucleic acid) has been codon-optimized for expression in cells of a subject, optionally wherein the subject is a non-human mammal or a human.

[0264] In specific embodiments, the nucleic acids described herein (e.g., non-naturally occurring nucleic acids) are mRNAs, and wherein thymidine (e.g., in the corresponding DNA sequence) is replaced by uracil or a functional analog thereof in the nucleic acid.

[0265] In specific embodiments, provided herein are nucleic acids described in Section 6, below. In specific embodiments, provided herein are codon-optimized nucleic acids described in Section 6, below. In specific embodiments, provided herein are non-naturally occurring nucleic acids described in Section 6, below. In some embodiments, provided herein are nucleic acids encoding mutants of VZV gE other than controls described in Section 6, below.

[0266] 5.4.25'-Cap structure

[0267] Without being bound by theory, it is expected that the 5'-cap structure of the polynucleotide participates in nuclear export and increases polynucleotide stability, and binds to mRNA cap binding protein (CBP), which is responsible for polynucleotide stability in the cell and causes translational competence by forming mature circular mRNA species through the association of CBP with poly-A binding protein. The 5'-cap structure further facilitates the removal of 5'-proximal introns during mRNA splicing. Therefore, in some embodiments, the nucleic acid molecules of the present disclosure comprise a 5'-cap structure.

[0268] Nucleic acid molecules can be capped at the 5' end by the cell's endogenous transcriptional machinery, thereby generating a 5'-ppp-5'-triphosphate linkage between the terminal guanosine cap residue of the polynucleotide and the sense nucleotide transcribed at the 5' end. This 5'-guanylate cap can then be methylated to generate an N7-methyl-guanylate residue. The ribose sugars of the terminal and / or anteterminal transcribed nucleotides at the 5' end of the polynucleotide can also be optionally 2'-O-methylated. 5'-decapping via hydrolysis and cleavage of the guanylate cap structure can target nucleic acid molecules, such as mRNA molecules, for degradation.

[0269] In some embodiments, the nucleic acid molecules of the present disclosure comprise one or more alterations to the native 5'-cap structure produced by endogenous processes. Without being bound by theory, modifications to the 5'-cap can increase the stability of the polynucleotide, increase the half-life of the polynucleotide, and increase the translation efficiency of the polynucleotide.

[0270] Exemplary changes to the natural 5'-cap structure include the production of a non-hydrolyzable cap structure to prevent decapping and thereby increase the half-life of the polynucleotide. In some embodiments, because the hydrolysis of the cap structure requires the cleavage of the 5'-ppp-5' phosphodiester linkage, in some embodiments, modified nucleotides can be used during the capping reaction. For example, in some embodiments, the vaccinia capping enzyme (Vaccinia Capping Enzyme) from New England Biolabs (Ipswich, Mass.) can be used for α-thioguanosine nucleotides to produce thiophosphate linkages in the 5'-ppp-5' cap according to the manufacturer's instructions. Additional modified guanosine nucleotides, such as α-methylphosphonic acid and selenophosphate nucleotides, can be used.

[0271] Additional exemplary alterations to the native 5'-cap structure include modifications at the 2' and / or 3' position of the capped guanosine triphosphate (GTP), replacement of the sugar ring oxygen (resulting in the oxygen of the carbocyclic ring) with a methylene moiety (CH2), modifications at the triphosphate bridge portion of the cap structure, or modifications at the nucleobase (G) portion.

[0272] Additional exemplary changes to the natural 5'-cap structure include, but are not limited to, 2'-O-methylation of the ribose sugar at the 5'-terminus of the polynucleotide and / or the 5'-terminal nucleotide on the sugar 2'-hydroxyl group (as described above). A variety of different 5'-cap structures can be used to produce the 5'-cap of a polynucleotide (such as an mRNA molecule). Additional exemplary 5'-cap structures that can be used in conjunction with the present disclosure also include those described in International Patent Publication Nos. WO 2008 / 127688, WO 2008 / 016473, and WO 2011 / 015347, the entire contents of each of which are incorporated herein by reference.

[0273] In various embodiments, the 5'-terminal cap may comprise a cap analog. Cap analogs are also referred to herein as synthetic cap analogs, chemical caps, chemical cap analogs, or structural or functional cap analogs, which differ in chemical structure from a natural (i.e., endogenous, wild-type, or physiological) 5'-cap while retaining cap function. Cap analogs can be synthesized and / or attached to polynucleotides chemically (i.e., non-enzymatically) or enzymatically.

[0274] For example, the anti-reverse cap analog (ARCA) cap contains two guanosines linked by a 5'-5'-triphosphate group, wherein one guanosine contains an N7-methyl group as well as a 3'-O-methyl group (i.e., N7, 3'-O-dimethyl-guanosine-5'-triphosphate-5'-guanosine, i.e., m7G-3'mppp-G, which can be equivalently referred to as 3'O-Me-m7G(5')ppp(5')G). The 3'-O atom of the other, unchanged guanosine is linked to the 5'-terminal nucleotide of the capped polynucleotide (e.g., mRNA). The N7- and 3'-O-methylated guanosines provide the terminal portion 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 the guanosine (i.e., N7, 2'-O-dimethyl-guanosine-5'-triphosphate-5'-guanosine, i.e., m7Gm-ppp-G).

[0275] In some embodiments, the cap analog can be a dinucleotide cap analog. As a non-limiting example, a dinucleotide cap analog can be modified with boranophosphate or phophoroselenoate at different phosphate positions, such as the dinucleotide cap analogs described in U.S. Patent No. 8,519,110, the entire contents of which are incorporated herein by reference in their entirety.

[0276] In some embodiments, the cap analog can be an N7-(4-chlorophenoxyethyl) substituted dinucleotide cap analog known in the art and / or described herein. Non-limiting examples of N7-(4-chlorophenoxyethyl) substituted dinucleotide cap analogs include N7-(4-chlorophenoxyethyl)-G(5')ppp(5')G and N7-(4-chlorophenoxyethyl)-m3'-OG(5')ppp(5')G cap analogs (see, for example, various cap analogs and methods for synthesizing cap analogs described in Kore et al. Bioorganic & Medicinal Chemistry 201321:4570-4574; the entire contents of the document are incorporated herein by reference). In other embodiments, the cap analog that can be used in conjunction with the nucleic acid molecules of the present disclosure is a 4-chloro / bromophenoxyethyl analog.

[0277] In various embodiments, the cap analog may comprise a guanosine analog. 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.

[0278] Without being bound by theory, it is expected that although cap analogs allow simultaneous capping of polynucleotides in in vitro transcription reactions, up to 20% of transcripts remain uncapped. This, along with the structural differences between cap analogs and the natural 5'-cap structure of polynucleotides produced by the cell's endogenous transcriptional machinery, may lead to reduced translational capacity and cellular stability.

[0279] Therefore, in some embodiments, the nucleic acid molecules of the present disclosure can also be capped after transcription using enzymes to produce a more authentic 5'-cap structure. As used herein, the phrase "more authentic" refers to a feature that closely reflects or mimics an endogenous or wild-type feature in structure or function. That is, a "more authentic" feature better represents endogenous, wild-type, natural or physiological cellular function and / or structure than a synthetic feature or analog of the prior art, or it outperforms the corresponding endogenous, wild-type, natural or physiological feature in one or more aspects. Non-limiting examples of more authentic 5'-cap structures that can be used in conjunction with the nucleic acid molecules of the present disclosure are structures that, in particular, have enhanced binding to cap-binding proteins, increased half-life, reduced sensitivity to 5'-endonucleases and / or reduced 5'-decapping compared to synthetic 5'-cap structures known in the art (or compared to wild-type, natural or physiological 5'-cap structures). For example, in some embodiments, a recombinant vaccinia virus capping enzyme and a recombinant 2'-O-methyltransferase can generate a classic 5'-5'-triphosphate linkage between the 5'-terminal nucleotide of a polynucleotide and a guanosine cap nucleotide, wherein the cap guanosine contains an N7-methylation and the 5'-terminal nucleotide of the polynucleotide contains a 2'-O-methyl group. This structure is referred to as the cap 1 structure. Compared to other 5' cap analog structures known in the art, for example, this cap results in higher translational capacity, cellular stability, and reduced activation of cellular proinflammatory cytokines. 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 comprises m 7 GpppAmpU.

[0280] Without being bound by theory, it is contemplated that the nucleic acid molecules of the present disclosure can be capped post-transcriptionally, and because this method is relatively efficient, nearly 100% of the nucleic acid molecules can be capped.

[0281] In some embodiments, provided herein are non-naturally occurring nucleic acids consisting of, consisting essentially of, or comprising: (1) the nucleotide sequence set forth 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 set forth in SEQ ID NO:63; except that all thymines (T) are substituted with uracils (U) or N1-methylpseudouridines, and / or the first nucleotide G is replaced with m 7 In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising: (1) the nucleotide sequence set forth 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 set forth in SEQ ID NO:63; except that all thymines (T) are substituted with uracils (U) or N1-methylpseudouridines, and / or the first nucleotide G is substituted with m 7 In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising the nucleotide sequence set forth in SEQ ID NO: 63, except that all thymines (T) are substituted with uracils (U) or N1-methylpseudouridines, and / or the first nucleotide G is substituted with m 7 In some embodiments, provided herein are nucleic acids consisting of, consisting essentially of, or comprising the nucleotide sequence shown in SEQ ID NO: 63, except that all thymines (T) are replaced by N1-methylpseudouridines, and the first nucleotide G is replaced by m 7 GpppAmpU replaced.

[0282] 5.4.3 Untranslated Region (UTR)

[0283] In some embodiments, the nucleic acid molecules of the present disclosure include one or more untranslated regions (UTRs). In some embodiments, the UTR is located upstream of the coding region in the nucleic acid molecule and is referred to as a 5'-UTR. In some embodiments, the UTR is located downstream of the coding region in the nucleic acid molecule and is referred to as a 3'-UTR. The sequence of the UTR may be homologous or heterologous to the sequence 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 gene origins. According to the present disclosure, any portion of the UTR in the nucleic acid molecule (including without any portion) may be codon optimized, and any portion may independently contain one or more different structures or chemical modifications before and / or after codon optimization.

[0284] In some embodiments, nucleic acid molecules (e.g., mRNA) of the present disclosure comprise UTRs and coding regions that are homologous to each other. In other embodiments, nucleic acid molecules (e.g., mRNA) of the present disclosure comprise UTRs and coding regions that are heterologous to each other. In some embodiments, to monitor the activity of UTR sequences, nucleic acid molecules comprising coding sequences of UTRs and detectable probes can be administered in vitro (e.g., in cell or tissue culture) or in vivo (e.g., to a subject), and the effects of the UTR sequences (e.g., modulating expression levels, cellular localization of the encoded product, or half-life of the encoded product) can be measured using methods known in the art.

[0285] In some embodiments, the UTR of the nucleic acid molecules (e.g., mRNA) of the present disclosure includes at least one translation enhancer element (TEE), which plays a role in increasing the amount of the polypeptide or protein produced by the nucleic acid molecules. In some embodiments, the TEE is located in 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 other embodiments, at least two TEEs are located at the 5'-UTR and 3'-UTR of the nucleic acid molecule, respectively. In some embodiments, the nucleic acid molecules (e.g., mRNA) of the present disclosure may include one or more copies of the TEE sequence or include more than one different TEE sequence. In some embodiments, the different TEE sequences present in the nucleic acid molecules of the present disclosure may be homologous or heterologous relative to each other.

[0286] Various TEE sequences are known in the art and can be used in conjunction with the present disclosure. For example, in some embodiments, the TEE can be an internal ribosome entry site (IRES), 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 (IRES) that can be used in conjunction with the present disclosure include, but are not limited to, U.S. Patent No. 7,468,275, U.S. Patent Publication No. 2007 / 0048776 and U.S. Patent Publication No. 2011 / 0124100, and the IRES described in International Patent Publication No. WO 2007 / 025008 and International Patent Publication No. WO 2001 / 055369, the contents of each of which are incorporated herein by reference in their entirety. In some embodiments, the TEE can be the TEE described in Supplementary Table 1 and Supplementary Table 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 herein by reference in their entirety.

[0287] Additional exemplary TEEs that may be used in conjunction with the present disclosure include, but are not limited to, U.S. Patent No. 6,310,197, U.S. Patent No. 6,849,405, U.S. Patent No. 7,456,273, U.S. Patent No. 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, U.S. Patent Publication No. 2009 / 0093049, International Patent Publication No. WO 2009 / 075886, International Patent Publication No. WO 2012 / 009644, and International Patent Publication No. WO 1999 / 024595, International Patent Publication No. WO 2007 / 025008, International Patent Publication No. WO TEE sequences described in EP 2001 / 055371, EP 2610341, EP 2610340, the contents of each of which are incorporated herein by reference in their entirety.

[0288] In various embodiments, nucleic acid molecules (e.g., mRNA) of the present disclosure include at least one UTR comprising 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 sequence in the UTR of nucleic acid molecules is a copy of the same TEE sequence. In other embodiments, at least two TEE sequences in the UTR of nucleic acid molecules have different TEE sequences. In some embodiments, a plurality of different TEE sequences are arranged in the UTR district of nucleic acid molecules with one or more repeat patterns. For illustration purposes only, repetitive pattern can be such as ABABAB, AABBAABBABB, ABCABCABC etc., wherein in these exemplary patterns, each capital letter (A, B or C) represents a different TEE sequence. In some embodiments, in the UTR of a nucleic acid molecule, at least two TEE sequences are continuous to each other (that is, without a spacer sequence therebetween). In other embodiments, at least two TEE sequences are separated by a spacer sequence. In some embodiments, UTR can include a TEE sequence-spacer sequence module, which is repeated in UTR at least once, at least twice, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times or more than 9 times. In any embodiment described in this paragraph, UTR can be 5'-UTR, 3'-UTR or both 5'-UTR and 3'-UTR of a nucleic acid molecule.

[0289] In some embodiments, the UTR of the nucleic acid molecule (e.g., mRNA) of the present disclosure includes at least one translation inhibition element, which acts to reduce the amount of the polypeptide or protein produced by the nucleic acid molecule. In some embodiments, the UTR of the nucleic acid molecule includes one or more miR sequences or fragments thereof (e.g., miR seed sequences (seed sequence)) identified by one or more microRNAs. In some embodiments, the UTR of the nucleic acid molecule includes one or more stem-loop structures that lower the translation activity of the nucleic acid molecule. Other mechanisms for suppressing the translation activity associated with the nucleic acid molecule are known in the art. In any embodiment described in this section, the UTR can be 5'-UTR, 3'-UTR, or both 5'-UTR and 3'-UTR of the nucleic acid molecule. Table 4 shows exemplary 5'-UTR and 3'-UTR sequences that can be used in conjunction with the present disclosure.

[0290] Table 4. Exemplary untranslated region (UTR) sequences.

[0291]

[0292]

[0293] In specific embodiments, the nucleic acid molecules of the present disclosure comprise a 5'-UTR selected from any one of SEQ ID NOs: 29-38. In specific embodiments, the nucleic acid molecules of the present disclosure comprise a 3'-UTR selected from any one of SEQ ID NOs: 39-46. In specific embodiments, the nucleic acid molecules of the present disclosure comprise 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 as described herein, such as any DNA coding sequence in Tables 1 to 4 or its equivalent RNA sequence. In specific embodiments, the nucleic acid molecules described in this paragraph may be in vitro transcribed RNA molecules.

[0294] Table 5 Exemplary DNA constructs

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316] A n = 120mer of A in the sequences in Table 5. More generally, for example, A n = 30 to 3000 polymers.

[0317] 5.4.4 Polyadenylation (Poly-A) Region

[0318] During natural RNA processing, long chains of adenosine nucleotides (poly-A regions) are typically added to messenger RNA (mRNA) molecules to increase the molecule's stability. Immediately after transcription, the 3'-end of the transcript is cleaved to release the 3'-hydroxyl group. Next, poly-A polymerase adds a series of adenosine nucleotides to the RNA. This process, called polyadenylation, adds poly-A regions between 100 and 250 residues in length. Without being bound by theory, it is expected that poly-A regions can confer multiple advantages to the nucleic acid molecules of the present disclosure.

[0319] Therefore, in some embodiments, the nucleic acid molecules (e.g., mRNA) of the present disclosure comprise a polyadenylation signal. In some embodiments, the nucleic acid molecules (e.g., mRNA) of the present disclosure comprise one or more polyadenylation (poly-A) regions. In some embodiments, the poly-A region is entirely composed of adenine nucleotides or functional analogs thereof. In some embodiments, the nucleic acid molecule comprises at least one poly-A region at its 3' end. In some embodiments, the nucleic acid molecule comprises at least one poly-A region at its 5' end. In some embodiments, the nucleic acid molecule comprises at least one poly-A region at its 5' end and at least one poly-A region at its 3' end.

[0320] According to the present disclosure, in different embodiments, the poly-A region may have different lengths. In particular, in some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 30 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 35 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 40 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 45 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 50 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 55 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 60 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 65 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 70 nucleotides. In some embodiments, the length of the poly-A region of the nucleic acid molecules of the present disclosure is at least 75 nucleotides. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 80 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 85 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 90 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 95 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 100 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 110 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 120 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 130 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 140 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 150 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 160 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 170 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 180 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 190 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 200 nucleotides in length.In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 225 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 250 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 275 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 300 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 350 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 400 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 450 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 500 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 600 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 700 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 800 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 900 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1000 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1100 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1200 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1300 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1400 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1500 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1600 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1700 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1800 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 1900 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 2000 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 2250 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 2500 nucleotides in length.In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 2750 nucleotides in length. In some embodiments, the poly-A region of the nucleic acid molecules of the present disclosure is at least 3000 nucleotides in length.

[0321] In some embodiments, the length of the poly-A region in a nucleic acid molecule can be selected based on the total length of the nucleic acid molecule or a portion thereof, such as the length of the co...

Claims

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

2. The protein of claim 1, wherein the mutant comprises the amino acid sequence of SEQ ID NO: 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 NO: 6, 8, 10, 12 or 3.

4. The protein of 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 of claim 1, wherein the mutant 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 of 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, 8, 10 or 12.

8. The protein of claim 1, wherein the mutant 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 of 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 of any one of claims 1 to 9, further comprising a VZV gE signal peptide.

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

18.

12. The protein of claim 11, wherein the signal peptide of the VZVgE 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 shown in SEQ ID NO:

18.

13. The protein of 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 of claim 13, wherein the heterologous signal peptide is a human IgE signal peptide.

15. The protein of claim 14, wherein the human IgE signal peptide comprises: (i) the amino acid sequence shown 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 shown in SEQ ID NO:

23.

16. The protein of claim 14, wherein the amino acid sequence of the human IgE signal peptide consists of the amino acid sequence shown in SEQ ID NO:

23.

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

23.

18. The protein of claim 14, comprising the amino acid sequence shown in SEQ ID NO:

59.

19. The protein of claim 14, wherein the amino acid sequence of the protein consists of the amino acid sequence shown in SEQ ID NO:

59.

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

21. The protein of claim 20, wherein the human tPA signal peptide comprises: (i) the amino acid sequence shown 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 shown in SEQ ID NO:

27.

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

55.

23. A fragment of the mature glycoprotein E (gE) of varicella-zoster virus (VZV), wherein the fragment comprises a truncation of at least one and at most 50, 49, 48, 47, 46, 45, 44, 43 or 42 amino acid residues from the C-terminus of the mature gE, optionally wherein the truncation is 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, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75 0, 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 the mature gE, or, optionally, the truncation is a truncation 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 truncation is a truncation of 14 or 37 amino acid residues from the C-terminus of the mature gE.

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

25. The fragment of claim 23 or 24, wherein the fragment comprises a truncation 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 of claim 25, wherein the fragment further comprises the amino acid residue substitution Y582G, and wherein amino acid residue position 582 is numbered according to the amino acid residue position of the full-length VZV gE protein.

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

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

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

30. The fragment of claim 29, wherein the fragment comprises the amino acid residue substitution S595A, and wherein amino acid residue position 595 is numbered according to the amino acid residue position of the full-length VSVgE.

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

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

33. A fragment as described in any one of claims 23 to 32, wherein the fragment comprises a truncation 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 1 amino acid residue from the C-terminus of the mature gE.

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

35. The fragment of claim 23, wherein the fragment comprises a truncation of the C-terminal 37 amino acid residues from the mature gE.

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

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

55.

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

39. A fusion protein comprising the fragment of 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 of claim 39, wherein the heterologous signal peptide is a human tPA signal peptide.

41. The fusion protein of claim 40, wherein the human tPA signal peptide comprises the amino acid sequence shown in SEQ ID NO:

27.

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

43. The fusion protein of claim 42, wherein the human IgE signal peptide comprises the amino acid sequence shown in SEQ ID NO:

23.

44. A fusion protein comprising a fragment of the mature glycoprotein (gE) of varicella-zoster virus (VZV) and a human IgE signal peptide, wherein the fragment comprises a truncation 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 the protein of any one of claims 1 to 22, the fragment of any one of claims 23 to 38, or the fusion protein of any one of claims 39 to 44.

46. ​​A nucleic acid as described in claim 45, wherein the nucleic acid comprises the nucleotide sequence shown in SEQ ID NO:7, 9, 11, 13, 4 or 5, or 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 shown in SEQ ID NO:7, 9, 11, 13, 4 or 5.

47. A nucleic acid encoding the protein of any one of claims 14 to 19 or the fusion protein of any one of claims 42 to 44, wherein the nucleotide sequence encoding the IgE signal peptide comprises: (a) the nucleotide sequence shown in SEQ ID NO:24, 25 or 26, 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 shown in SEQ ID NO:24, 25 or 26.

48. A nucleic acid encoding the protein of any one of claims 10 to 12, wherein the nucleotide sequence of the signal peptide comprises: (a) the nucleotide sequence shown in SEQ ID NO: 19, 20, 21 or 22, 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 shown in SEQ ID NO: 19, 20, 21 or 22.

49. A nucleic acid encoding the protein of claim 20 or 21 or the fusion protein of claim 40 or 41, wherein the nucleotide sequence encoding the human tPA signal peptide comprises: (a) the nucleotide sequence shown in SEQ ID NO:28, 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 shown in SEQ ID NO:

28.

50. A non-naturally occurring nucleic acid comprising a nucleotide sequence encoding the protein of any one of claims 1 to 22, the fragment of any one of claims 23 to 38, or the fusion protein of any one of claims 39 to 44.

51. The non-naturally occurring nucleic acid of claim 50, wherein the encoding nucleotide sequence has been codon-optimized for expression in a cell of a subject, optionally wherein the subject is a non-human mammal or a human.

52. The non-naturally occurring nucleic acid of claim 50 or 51, comprising: (a) the nucleotide sequence shown 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 shown in SEQ ID NO:7, 9, 11, 13, 4 or 5.

53. A non-naturally occurring nucleic acid encoding the protein of any one of claims 14 to 19 or the fusion protein of any one of claims 42 to 44, wherein the nucleotide sequence encoding the IgE signal peptide comprises: (a) the nucleotide sequence shown in SEQ ID NO:24, 25 or 26, 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 shown in SEQ ID NO:24, 25 or 26.

54. A non-naturally occurring nucleic acid encoding the protein of any one of claims 10 to 12, wherein the nucleotide sequence of the signal peptide of the full-length VSVgE protein comprises: (a) the nucleotide sequence shown in SEQ ID NO: 19, 20, 21 or 22, 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 shown in SEQ ID NO: 19, 20, 21 or 22.

55. A non-naturally occurring nucleic acid encoding the protein of claim 20 or 21 or the fusion protein of claim 40 or 41, wherein the nucleotide sequence encoding the human tPA signal peptide comprises: (a) the nucleotide sequence shown in SEQ ID NO:28, 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 shown in SEQ ID NO:

28.

56. A non-naturally occurring nucleic acid comprising the nucleotide sequence of SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4 or 5.

57. The non-naturally occurring nucleic acid of claim 56, which consists of the nucleotide sequence of SEQ ID NO: 63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4 or 5.

58. The non-naturally occurring nucleic acid of any one of claims 50 to 55, wherein the nucleic acid consists of, consists essentially of, or comprises: (1) the nucleotide sequence shown in SEQ ID NO:63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5; 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 shown in SEQ ID NO:63, 7, 51, 60, 62, 62, 64, 9, 11, 13, 52, 53, 54, 58, 49, 50, 4, or 5.

59. The non-naturally occurring nucleic acid of any one of claims 50 to 58, The non-naturally occurring nucleic acid further comprises a 5' untranslated region (5'-UTR), wherein the 5'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 29-38; and / or, The non-naturally occurring nucleic acid further comprises a 3' untranslated region (3'-UTR), wherein the 3'-UTR comprises the nucleotide sequence shown in any one of SEQ ID NOs: 39-46, Optionally, the 3'-UTR further comprises a poly-A tail or a polyadenylation signal.

60. The non-naturally occurring nucleic acid of any one of claims 50 to 59, wherein the nucleic acid is DNA.

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

62. The non-naturally occurring nucleic acid of any one of 50 to 59, wherein the nucleic acid is an mRNA, and wherein thymine is replaced by uracil or a functional analog thereof in the nucleic acid.

63. The non-naturally occurring nucleic acid of claim 62, wherein the nucleic acid comprises one or more functional nucleotide analogs selected from pseudouridine, 1-methyl-pseudouridine, and 5-methylcytosine.

64. The non-naturally occurring nucleic acid of claim 62, wherein the nucleic acid consists of, consists essentially of, or comprises: (1) the nucleotide sequence set forth 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 set forth in SEQ ID NO:63; except that all thymines (T) are replaced with uracils (U) or N1-methylpseudouridines, and / or the first nucleotide G is replaced with m 7 Replaced by GpppAmpU.

65. The non-naturally occurring nucleic acid of claim 62, wherein the nucleic acid consists of, consists essentially of, or comprises the nucleotide sequence set forth in SEQ ID NO:63, except that all thymines (T) are replaced with uracils (U) or N1-methylpseudouridines, and / or the first nucleotide G is replaced with m 7 Replaced by GpppAmpU.

66. The non-naturally occurring nucleic acid of claim 62, wherein the nucleic acid consists of, consists essentially of, or comprises the nucleotide sequence set forth in SEQ ID NO:63, except that all thymines (T) are replaced with N1-methylpseudouridines, and the first nucleotide G is replaced with m 7 Replaced by GpppAmpU.

67. A vector comprising the nucleic acid of any one of claims 45 to 49 or the non-naturally occurring nucleic acid of any one of claims 50 to 66; preferably, the vector is an IVT (in vitro transcription) plasmid.

68. A host cell comprising the nucleic acid of any one of claims 45 to 49, the non-naturally occurring nucleic acid of any one of claims 50 to 66, or the vector of claim 67.

69. The host cell of claim 68, wherein the host cell is in vitro, ex vivo or isolated.

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

71. A pharmaceutical composition comprising the nucleic acid of any one of claims 45 to 49, the non-naturally occurring nucleic acid of any one of claims 50 to 66, or the vector of claim 67.

72. A pharmaceutical composition comprising a non-naturally occurring nucleic acid as described in any one of claims 50 to 66 and at least a first lipid, optionally wherein 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 the compounds listed in Table 02-1; Or a compound according to formula 03-I; or the compounds listed in Table 03-1; Or a compound according to formula 04-I; Or the compounds listed in Table 04-1.

73. A pharmaceutical composition as described in claim 72, further comprising a second lipid, optionally, wherein the second lipid is a compound according to Formula 05-I.

74. The pharmaceutical composition of claim 72 or 73, formulated as lipid nanoparticles encapsulating the nucleic acid in a lipid shell.

75. A pharmaceutical composition as described in any one of claims 70 to 74, wherein the composition is a vaccine.

76. A method for controlling, preventing or treating a disease or condition caused by VZV or infection with VZV in a subject, the method comprising administering to the subject a therapeutically effective amount of a protein as described in any one of claims 1 to 22, a fragment as described in any one of claims 23 to 38, a fusion protein as described in any one of claims 39 to 44, a nucleic acid as described in any one of claims 45 to 49, a non-naturally occurring nucleic acid as described in any one of claims 50 to 66, a vector as described in claim 67, or a pharmaceutical composition as described in any one of claims 70 to 75.

77. The method of claim 76, wherein the method is used to prevent a disease or condition caused by VZV or by infection with VZV in the subject.

78. The method of claim 76 or 77, wherein an immune response against the VZV is elicited in the subject.

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

80. The method of claim 78 or 79, wherein the immune response comprises an increase in the proportion of lymphocytes expressing cytokines.

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

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

83. The method of any one of claims 78 to 82, wherein the immune response comprises the production of antibodies that specifically bind to VZV gE.

84. The method of any one of claims 76 to 83, wherein the disease or condition caused by VZV is (a) Chickenpox 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 of any one of claims 76 to 84, wherein the subject is a human.

86. The method of claim 85, wherein the human is a human adult.

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

88. The method of claim 85, wherein the human is an elderly human.

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 non-naturally occurring nucleic acid 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 of controlling, preventing or treating a disease or condition caused by VZV or infection with VZV in a subject.

90. The protein, fragment, fusion protein, nucleic acid, non-naturally occurring nucleic acid, vector or pharmaceutical composition for use according to claim 89, wherein the subject is a human, optionally a human adult or elderly.

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 non-naturally occurring nucleic acid 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 the manufacture of a medicament for controlling, preventing or treating a disease or condition caused by VZV or infection with VZV in a subject.

92. The use of claim 91, wherein the subject is a human, optionally an adult or elderly person.

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