Nucleic acid compositions for the generation of a ZIKA virus-like particle

Abstract

Disclosed herein are nucleic acid compositions for delivering polynucleotides of interest.

Description

NUCLEIC ACID COMPOSITIONS FOR THE GENERATION OF A ZIKAVIRUS-LIKE PARTICLECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 578,335, filed on August 23, 2023, which is incorporated herein by reference in its entirety.SEQUENCE LISTING

[0002] This application contains a Sequence Listing which has been submitted in XML format and is hereby incorporated by reference in its entirety. The XML copy, created on August 20, 2024, is named 63070_704_601_SL.xml, and is 3,010,207 bytes in size.BACKGROUND OF THE INVENTION

[0002] The cellular delivery of various biologically active compounds, such as an antigen and a protein of interest, is of high value both for basic research and industrial applications. However, it is usually compromised by two limitations, one being high toxicity to normal tissues, the other being low delivery efficiency, given that living cells are highly restricted by the complex membrane systems of the cell. Special difficulties have been seen in delivery efforts with the central nervous system (CNS) and ocular system as a target.SUMMARY OF THE INVENTION

[0003] In one aspect, provided herein is a zika virus-like particle, comprising: a polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the polynucleotide does not encode one or more viral non-structural proteins and one or more viral structural proteins. In some embodiments, the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, and zika virus NS5 or a derivative of the zika virus NS5; and wherein the one or more viral structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, and zika virus viral envelope protein (E) or a derivative of the zika virus E. The polynucleotide may comprise a sequence encoding a protein of interest.

[0004] In one aspect, provided herein is a method of producing the zika virus-like particle described herein, the method comprising introducing into a host cell: the polynucleotide encoding the zika virus 5’ UTR and zika virus 3’ UTR, e.g., as described herein, a second polynucleotide encoding the one or more viral non-structural proteins, and a third polynucleotide encoding the one or more viral structural proteins. In some embodiments, each of the second polynucleotide and the third polynucleotide independently encode (i) a 5’ UTR that is not a 5’UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus, and (ii) a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus.

[0005] In one aspect, provided herein is a zika virus-like particle, comprising: a polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the polynucleotide does not encode one or more viral non-structural proteins and one or more viral structural proteins. In some embodiments, the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS 1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, and zika virus NS5 or a derivative of the zika virus NS5; and wherein the one or more viral structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, and zika virus viral envelope protein (E) or a derivative of the zika virus E. In some embodiments, the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, or zika virus NS5 or a derivative of the zika virus NS5, or a combination of two or more thereof. In some embodiments, the polynucleotide encodes a zika virus NS4a comprising a sequence at least 90% identical to SEQ ID NO: 2929 or 2946. In some embodiments, the polynucleotide encodes a zika virus NS4a comprising a pTMS3. In some embodiments, the polynucleotide does not encode a zika virus NS4a comprising a pTMSl and a pTMS2. In some embodiments, the polynucleotide does not encode a full length zika virus NS4a. In some embodiments, the full length zika virus NS4a comprises a sequence at least 90% identical to SEQ ID NO: 10 or 2942. In some embodiments, the polynucleotide encodes a NS4b comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948. In some embodiments, the polynucleotide encodes a NS4b comprising a N-terminal disordered region. In some embodiments, the polynucleotide does not encode a zika virus NS4b comprising a pTMSl, a pTMS2, a pTMS3, a pTMS4, and a pTMS5. In some embodiments, the polynucleotide does not encode a full length zika virus NS4b. In some embodiments, the full length zika virus NS4b comprises a sequence at least 90% identical to SEQ ID NO: 12 or 2944. In some embodiments, the polynucleotide encodes a zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947. In some embodiments, the one or more structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, zika virus viral envelope protein (E) or a derivative of the zika virus E, or a combination thereof. In some embodiments, the polynucleotide encodes a zika virus capsid protein (C) comprising a sequenceat least 90% identical to any one of SEQ ID NOS: 2908-2917. In some embodiments, the polynucleotide does not encode a full length zika virus C. In some embodiments, the full length zika virus C comprises a sequence at least 90% identical to SEQ ID NO: 3 or 2901. In some embodiments, the zika virus-like particle comprises the one or more viral non-structural proteins and the one or more viral structural proteins, optionally wherein the one or more structural proteins are assembled into a zika virus-like shell, and further optionally wherein the zika viruslike particle is not replication competent. The polynucleotide may comprise a sequence encoding a protein of interest.

[0006] In one aspect, provided herein is a method of producing the zika virus-like particle described herein, the method comprising introducing into a host cell: the polynucleotide encoding the zika virus 5’ UTR and the zika virus 3’ UTR, a second polynucleotide encoding the one or more viral non-structural proteins, and a third polynucleotide encoding the one or more viral structural proteins. In some embodiments, each of the second polynucleotide and the third polynucleotide independently comprise a 5’ UTR that is not a zika virus 5’ UTR, and a 3’ UTR that is not a zika virus 3’ UTR. In some embodiments, the zika virus-like particle described herein, or the method described herein, wherein the zika virus 5’ UTR comprises a sequence at least 90% identical to any one of SEQ ID NOS: 1, 324-633, or the 5’ UTR of the zika nucleotide sequences of Table IB; and wherein the zika virus 3’ UTR comprises a sequence at least 90% identical to any one of SEQ ID NOS: 2, 14-323, or the 3’ UTR of the zika nucleotide sequences of Table IB.

[0007] In one aspect, provided herein is a composition comprising: (a) a first polynucleotide encoding a zika virus structural protein, (b) a second polynucleotide encoding a zika virus non- structural (NS) protein, and (c) a third polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the first polynucleotide and the second polynucleotide independently do not encode the zika virus 5’ UTR or the zika virus 3’ UTR. The third polynucleotide may comprise a sequence encoding a protein of interest. In some embodiments, the first polynucleotide encoding the zika virus structural protein comprises: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C; (d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; (e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E, or (f) a combination of two or more selected from (a) to (e). In some embodiments, the 5’ UTRof the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919. In some embodiments, the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921. In some embodiments, the second polynucleotide encoding a zika virus non-structural (NS) protein comprises: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1; (d) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A; (e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A; (f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B; (g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3; (h) a polynucleotide encoding a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A; (i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K; (j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B; (k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5; or (1) a combination of two or more selected from (a) to (k). In some embodiments, the second polypeptide further comprises a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E. In some embodiments, the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919. In some embodiments, the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921. In some embodiments, the third polynucleotide encoding a zika virus 5’ UTR of Table IB and a zika virus 3’ UTR of Table IB comprises: (a) a polynucleotide encoding a zika virus 5’ UTR of the zika nucleotide sequences of Table IB; (b) a polynucleotide encoding a zika virus 3’ UTR of the zika nucleotide sequences of Table IB; (c) a polynucleotide of interest; (d) a non-structural protein, or a derivative of a non-structural protein; (e) a structural protein, or a derivative of a structural protein; or (f) a combination of two or more selected from (a) to (e). In some embodiments, the non-structural protein comprises a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A, a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, and a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B. In someembodiments, the non- structural protein comprises a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A comprising a sequence at least 90% identical to SEQ ID NO: 2929 or 2946, a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947, and a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948. In some embodiments, the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C. In some embodiments, the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C comprising a sequence at least 90% identical to SEQ ID NOS: 2908- 2917.

[0008] In one aspect, provided herein is a method of producing a zika virus like particle comprising introducing into a host cell the composition described herein.

[0009] In one aspect, provided herein is a structural protein plasmid comprising: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C; (d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; and (e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E; wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919, and wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0010] In one aspect, provided herein is a non-structural protein plasmid comprising: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1; (d) a polynucleotide encoding a zika virus non-structural protein 2A (NS2A) or the derivative of the zika virus NS2A; (e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A; (f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B; (g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3; (h) a polynucleotide encoding a zika virus non-structural protein 4A (NS4A) or thederivative of the zika virus NS4A; (i) a polynucleotide encoding a zika virus non- structural protein 2K (2K) or the derivative of the zika virus 2K; (j) a polynucleotide encoding a zika virus non- structural protein NS4B (NS4B) or the derivative of the zika virus NS4B; (k) a polynucleotide encoding a zika virus non- structural protein NS5 (NS5) or the derivative of the zika virus NS5; (1) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E; or (m) a combination of two or more selected from (a) to (1); wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919, and wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0011] In one aspect, provided herein is a polynucleotide of interest plasmid comprising: (a) a polynucleotide encoding a zika virus 5’ UTR of the zika nucleotide sequences of Table IB; (b) a polynucleotide encoding a zika virus 3’ UTR of the zika nucleotide sequences of Table IB; (c) a polynucleotide of interest; (d) a non-structural protein, or a derivative of a non-structural protein; (e) a structural protein, or a derivative of a structural protein; or (f) a combination of two or more selected from (a) to (e); wherein the non-structural protein comprises a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, and a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, and wherein the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C. The polynucleotide may comprise a sequence encoding a protein of interest. In some embodiments, the non-structural protein comprises a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A comprising a sequence at least 90% identical to SEQ ID NO: 2929 or 2946, the zika virus non- structural protein 2K (2K) or the derivative of the zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947, and the zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948. In some embodiments, the structural protein comprises a zika virus capsid protein (C), or the derivative of a zika virus C comprising a sequence at least 90% identical to SEQ ID NOS: 2908-2917.

[0012] In one aspect, provided herein is a method for producing a zika virus-like particle, comprising introducing into a host cell: (a) the structural protein plasmid described herein, (b) the non-structural protein plasmid described herein, and (c) the polynucleotide of interest plasmid described herein.

[0013] In one aspect, provided herein is a method of treating a disease or condition, comprising administering to a subject in need thereof the zika virus-like particle described herein or the zika virus-like particle produced by the method described herein.

[0014] In one aspect, provided herein is a method of triggering or boosting an immune response, the method comprising administering to a subject in need thereof an effective amount of the zika virus-like particle described herein, or the zika virus-like particle produced by the method described herein.

[0015] A vaccine composition comprising the zika virus-like particle described herein, or the zika virus-like particle produced by the method described herein.Brief Description of the Drawings

[0016] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0017] FIG. 1 shows a plasmid construct comprising a hammerhead ribozyme of a zika virus; (ii) a 5’ Untranslated region (UTR) of the zika virus; (iii) a C-partial of the zika virus; (iv) a 3’ UTR of the zika virus; (v) a hepatitis delta virus (HDV) ribozyme of the zika virus, and two reporter genes (eGFP) and Nluc.

[0018] FIGS. 2A-2E are example pREP constructs having different lengths of partial capsid gene: 15 bp (FIG. 2A), 30 bp (FIG. 2B), 45 bp (FIG. 2C), 60 bp (FIG. 2D), and 75 bp (FIG. 2E).

[0019] FIG. 3 shows pREP RNA encapsulation in all plasmids tested (FIGS. 2A-2E, C-partial with 15, 30, 45, 60 and 75 bp, respectively).

[0020] FIG. 4 shows Nanoluc quantification in Vero cells after contact with viral vector (pREP, e.g., of FIGS. 2A-2E + ZIKV) carrying a gene encoding 5, 10, 15, 20 or 25 amino acids of capsid C, derived from RNA transfection, compared to ZIKV (no pREP, ZIKV only infection). A higher level of NanoLuc expression is demonstrated in viral vectors of all capsid sequence sizes compared to the control group. These results indicate that ZIKVV is able to encapsulate pREP RNA with C-partial (15, 30, 45, 60 and 75 bp) and deliver NanoLuc, confirmed by the expression of the reporter gene in the target cells.

[0021] FIG. 5 shows a plasmid construct comprising sequences of DNA polymerase promoter, RNA polymerase promoter, Xenopus globin 5’ UTR, internal ribosome entry site (IRES), Xenopus globin 3’ UTR and encoding Capsid, Membrane, Envelope and Nluc luciferase.

[0022] FIG. 6 shows a plasmid construct comprising sequences of DNA polymerase promoter, RNA polymerase promoter, tobacco etch virus (TEV) 5’ UTRs, Xenopus globin 3’ UTR, 2 A self-cleaving peptide (P2A), Nluc luciferase, Envelope-partial protein (E), non-structural protein 1 (NS1), non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, non- structural protein 2B (NS2B), non-structural protein 3 (NS3), non-structural protein 4A (NS4A), non-structural protein 2K (2K), non-structural protein NS4B (NS4B) and non-structural protein NS5 (NS5).

[0023] FIG. 7A and FIG. 7B show Nluc activities observed in the cell types Vero and EEK-293 after transfection with pREP plasmid (e.g., as shown in FIGS. 1, 2A-2E, 9, 10A-10F), structural protein plasmid (e.g., as shown in FIGS. 5, 8, 11), and non-structural protein plasmid (e.g., as shown in FIGS. 6, 12).

[0024] FIG. 8 shows a plasmid construct comprising sequences of DNA polymerase promoter, RNA polymerase promoter, Xenopus globin 5’ UTR, internal ribosome entry site (IRES), Xenopus globin 3’ UTR and encoding Capsid, Membrane, and Envelope.

[0025] FIG. 9 shows a plasmid construct comprising a eukaryotic enhancer, a eukaryotic promoter, a T7 promoter, a hammerhead ribozyme of a zika virus, a 5’ Untranslated region (UTR) of the zika virus, a C (capsid) protein partial of the zika virus, NS4a partial protein, 2K, and NS4b partial protein, two reporter genes (eGFP) and Nanoluciferase, a 3’ UTR of the zika virus, a hepatitis delta virus (HDV) ribozyme of the zika virus, and a bGH poly(A) signal.

[0026] FIGS. 10A-10F are example constructs of FIG. 9 having different lengths of partial C (capsid) gene: 0 bp (FIG. 10A), 15 bp (FIG. 10B), 30 bp (FIG. 10C), 45 bp (FIG. 10D), 60 bp (FIG. 10E), and 75 bp (FIG. 10F).

[0027] FIG. 11 shows a plasmid construct comprising sequences of CMV enhancer and CMV promoter, T7 promoter, Xenopus globin 5’ UTR, encoding Capsid, Membrane, Envelope and Nluc luciferase, internal ribosome entry site (IRES), Xenopus globin 3’ UTR and bGH poly(A) signal.

[0028] FIG. 12 shows a plasmid construct comprising sequences of CMV enhancer and promoter, T7 promoter, tobacco etch virus (TEV) 5’ UTRs, Nluc luciferase, 2 A self-cleaving peptide (P2A), Envelope-partial protein (E), non-structural protein 1 (NS1), non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, non-structural protein 2B (NS2B), non-structural protein 3 (NS3), non-structural protein 4A (NS4A), non-structural protein 2K (2K), non-structural protein NS4B (NS4B) and non-structural protein NS5 (NS5), Xenopus globin 3’ UTR, and bGH poly(A) signal.

[0029] FIGS. 13A and 13B show the schematic representation of viral vector production. FIG. 13A shows simplified construction of plasmids. From top to bottom, expression of structural proteins (pStructural) (simplified FIG. 11), helper plasmid for non- structural proteins (pNSP) (simplified FIG. 12), and plasmid for translation of mRNA containing reporter genes of interest (pREP) (simplified FIG. 9). FIG. 13B shows a schematic of the co-transfection of the three plasmids in cell culture and the production of the viral vector. In other examples, the reporter or other gene of interest is only present in the pREP plasmid, and not present in the pStructural or pNSP plasmids.

[0030] FIGS. 14A and 14B show the RNA quantification after collecting the supernatant at 24, 48 and 72 hours after transfection (e.g., using the method shown in FIG. 13B), an RT-qPCR analysis was performed to quantify the secreted material RNA, and the result presented highlights the best RT-qPCR analysis time which was 72 hours after transfection in both cells. FIG. 14A shows co-transfection of plasmids into HEK-293T cells; FIG. 14B shows cotransfection of plasmids into Vero cells.

[0031] FIGS. 15A and 15B show the expression of the Nanoluciferase protein evaluation after the transfection with pREP (e.g., as shown in FIG. 9, 10A-10F), pStructural (e.g., as shown in FIG. 11), and / or pNSP (e.g., as shown in FIG. 12) was used to transduce Vero cells. Supernatant was collected 24-, 48-, and 72-hours post-transfection. FIG. 15A shows Nluc activity from supernatant collected after transfection into Vero cells. FIG. 15B shows Nluc activity from supernatant after transfection into HEK-293T cells. *p<0.05, **p<0.01, ***p<0.0001 according to one-way ANOVA with bonferroni correction.

[0032] FIG. 16 shows the expression of the Nanoluciferase protein by measuring Nluc activity after the viral vectors (e.g., as shown in FIGS. 9 or 10A-10F, 11, and 12) were used to transduce Vero cells. Supernatant collected 72- and 96-hours post-transfection into HEK 293T cells. *p<0.05, **p<0.01, ***p<0.0001 according to one-way ANOVA with bonferroni correction.

[0033] FIG. 17 shows the Western blot of sZIKV-V obtained at cell culture media after 96 hours of co-transfection of the three plasmids (Lane 2) (e.g., as shown in FIGS. 9 or 10A-F, 11, and 12), and oncolytic Zika virus used as positive control (Lane 3). An anti-Zika envelope protein antibody was used for protein detection, and a protein mass Ladder the determine the mass of the Envelope protein (Lane 1).

[0034] FIG. 18 shows the expression analysis of the NanoLuciferase enzyme by measuring Nluc activity after delivery of the viral vectors (e.g., as shown in FIGS. 9 or 10A-F, 11, and 12) to verify in vitro tropism in Vero, MCF7 and CHLA-06 strains. *p<0.05, **p<0.01, ***p<0.0001 according to one-way ANOVA with bonferroni correction.

[0035] FIG. 19 shows the Heat map of the multiplex immunoassay indicating the pro- inflammatory, chemokine, and growth factor responses in animals administered with sZIKV-V (e.g., the viral vectors of FIGS. 9 or 10A-F, 11, and 12) or wild-type ZIKV. The analysis was performed on blood collected 0- (before, control), 3-, 7- and 14-days post-injection. Higher levels of pro-inflammatory molecules were observed in animals treated with the wild-type zika virus as compared to animals treated with the sZIKV-V, e.g., INF gamma (3 days, 7 days), TNF alpha (14 days), IL1 beta (3 days), IL6 (3 days), IL7 (3 days, 14 days), IL-7R alpha (3 days), IL9 (3, 14 days), IL-12p70 (3, 14 days), IL-15 (3, 14 days), and IL-18 (14 days). Higher levels of chemokines were observed in animals treated with the wild-type zika virus as compared to animals treated with the sZIKV-V, e.g., CXCL1 (14 days), CXCL10 (14 days), CCL3 (3, 14 days), CCL4 (3 days), CCL5 (3, 14 days), and CCL7 (14 days). Higher levels of growth factors were observed in animals treated with wild-type zika virus as compared to treated with sZIKV-V, e.g., BAFF, G-CSF and VEGF-A at 14 days.

[0036] FIG. 20 shows the immunofluorescence of histological brain sections (from the cortex and hippocampus regions) of various groups: control animals (Control) that did not receive any virus or viral vector, animals administered a dose of the synthetic viral vector (sZIKV-V) carrying pREP-EGFP-NanoLuc (e.g., the viral vectors of FIGS. 9 or 10A-F, 11, and 12), animals inoculated with a dose of wild type Zika virus (ZIKV). The nuclear staining is shown in blue with DAPI, the GFP protein is labeled in red with AF647, and the NS2 protein is depicted in green with AF488. Tissue samples were collected at 0-, 3-, 7-, and 14-days post-administration of the compounds.DETAILED DESCRIPTION OF THE INVENTION

[0037] Various nucleic acid compositions are provided herein comprising a polynucleotide of interest. The nucleic acid compositions may be useful for delivering the polynucleotide of interest to a target cell. The target cell may be a cell of the central nervous system or ocular system. The target cell may be present in a subject, where the nucleic acid compositions deliver the polynucleotide of interest to the target cell in the subject. As a non-limiting example, polynucleotides of interest may be contained within a nucleic acid composition referred to as a pREP plasmid herein. Certain nucleic acid compositions comprise an polynucleotide of interest up to about 6,000 or 10,000 nucleotides in length, e.g., about 5 nucleotides to about 6,000 nucleotides. The nucleic acid compositions may comprise a 5’ Untranslated region (UTR) of a zika virus and a 3’ UTR of the zika virus, and optionally do not comprise a polynucleotide encoding one or more nonstructural proteins of a virus, and further optionally do not comprise a polynucleotide encoding one or more structural proteins of a virus. Certain nucleic acidcompositions encode one or more structural proteins, e.g, as exemplified by pStructural constructs herein. Certain nucleic acid compositions encode one or more nonstructural proteins, e.g., as exemplified by pNSP constructs herein.

[0038] In one aspect, a zika virus-like particle is generated from three polynucleotides. The three polynucleotides may be present on three separate plasmids. As a non-limiting example, a pREP, pStructural, and pNSP construct herein. In some embodiments, a first polynucleotide encodes for one or more structural proteins of a zika virus. The structural proteins may be wild-type, functional fragments of wild-type proteins, or proteins having at least 80% sequence identity to wild-type proteins. The first polynucleotide may be a structural protein plasmid (e.g., pStructural). In some embodiments, a second polynucleotide encodes one or more nonstructural proteins. In some embodiments, this second polynucleotide is a helper plasmid. In some embodiments, the helper plasmid improves virus like particle assembly efficiency. The second polynucleotide may be a non-structural protein plasmid (e.g., pNSP). In some embodiments, a third polynucleotide comprises a polynucleotide of interest, or a polynucleotide of interest nucleic acid, that encodes a nucleic acid or peptide of interest, e.g., a therapeutic nucleic acid or peptide. The third polynucleotide may be a polynucleotide of interest plasmid (e.g., pREP). In some embodiments, the third polynucleotide is encapsulated by the zika virus-like particle. In an example embodiment, the zika virus-like particle comprises the one or more structural proteins of a zika virus, and the third polynucleotide, wherein the third polynucleotide is encapsulated by the zika virus-like particle. In some embodiments, the zika virus-like particle is delivered to the CNS. In some embodiments, the zika virus-like particle is delivered to the ocular system. In some embodiments, the first polynucleotide and the second polynucleotide are present on separate nucleic acids. In some embodiments, the first polynucleotide and the second polynucleotide are present on the same nucleic acid. In some embodiments, the structural protein plasmid and the non-structural protein plasmid are present on separate nucleic acids. In some embodiments, the structural protein plasmid and the non-structural protein plasmid are present on the same nucleic acid.

[0039] In one aspect, provided herein is a nucleic acid composition comprising a polynucleotide encoding a 5’ Untranslated region (UTR) of a zika virus; 3’ UTR of the zika virus; and a polynucleotide of interest. In one aspect, provided herein is a nucleic acid composition comprising (i) a polynucleotide encoding a ribozyme (ii) 5’ Untranslated region (UTR) of a zika virus; (iii) 3’ UTR of the zika virus; and (iv) a polynucleotide of interest. In some embodiments, the ribozyme is a hammerhead ribozyme or a hepatitis delta virus (HDV) ribozyme. In some embodiments, the nucleic acid further comprises a polynucleotide encoding a second ribozyme.In some specific embodiments, the second ribozyme is a hammerhead ribozyme or a hepatitis delta virus (HDV) ribozyme. In some embodiments, the polynucleotide encoding the ribozyme encodes a hammerhead ribozyme and the polynucleotide encoding the second ribozyme encodes an HDV ribozyme. In some embodiments, the nucleic acid composition further comprises a C- partial. For example, the C-partial is a derivative of C, where the derivative encodes a truncation (e.g., about or less than about 30, 25, 20, 15, 10, or 5 amino acids) of C. In some embodiments, the nucleic acid is a polynucleotide of a polynucleotide of interest plasmid. The nucleic acid composition may be packaged within a zika virus-like particle. The nucleic acid composition may comprise a promoter, such as a eukaryotic promoter. The nucleic acid composition may encode part of a NS4a. The nucleic acid composition may encode part of a NS4b. The nucleic acid composition may comprise a polyA region. Non-limiting example nucleic acid compositions include those shown in FIGS. 1, 2A-2E, 9 and 10A-10F.

[0040] In one aspect, provided herein is a nucleic acid composition comprising a sequence encoding one of the structural proteins from zika virus or its derivative with a polynucleotide of interest. In a non-limiting example, the nucleic acid composition comprises a derivative of zika capsid protein (C), where the derivative encodes a truncation (e.g., about or less than about 30, 25, 20, 15, 10, or 5 amino acids) of C. The nucleic acid composition may be a polynucleotide of a structural protein plasmid. The nucleic acid composition may be part of a polynucleotide of interest plasmid. Non-limiting example nucleic acid compositions include those shown in FIGS. 1, 2A-2E, 5, 8, 9, 10A-10F and 11.

[0041] Non-limiting example nucleic acid compositions herein may comprise one or more sequences from Tables 1 A-1F, Table 2, Table 3, and Table 4.

[0042] Further provided herein is a zika virus-like particle that comprises a zika virus capsid protein or a derivative thereof, or all three structural proteins from zika virus or their derivative (a zika virus capsid protein (C) or the derivative of the zika virus C, a polynucleotide encoding a zika virus viral membrane protein (prM / M) or the derivative of the zika virus prM / M, a polynucleotide encoding a zika virus viral envelope protein (E) or the derivative of the zika virus E). Non-limiting example nucleic acid compositions encoding structural proteins from zika virus that may be assembled into the zika virus-like particle include those shown in FIGS. 5, 8, and 11. Non-limiting example nucleic acid compositions encoding non- structural proteins from zika virus that may be assembled into the zika virus-like particle include those shown in FIGS. 6 and 12. Further provided herein is a method of using such zika virus-like particle to deliver polynucleotides of interest to a target cell. Non-limiting example nucleic acid compositions encoding a polynucleotide of interest that may be encapsulated into a zika virus-like particleinclude those shown in FIGS. 1, 2A-2E, 9, and 10A-10F. In preferred embodiments, such zika virus-like particle effectively crosses the blood brain barrier, resulting in a tropism targeting central nervous system (CNS). Accordingly, in preferred embodiments, such zika virus-like particle is used to deliver polynucleotides of interest to CNS. In preferred embodiments, the zika virus-like particle targets the ocular system.

[0043] Before the present methods and compositions are described, it is to be understood that this disclosure is not limited to a particular method or composition described, and as such may vary. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. It is understood that the present disclosure supersedes any disclosure of an incorporated publication to the extent there is a contradiction.

[0044] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

[0045] Where a range of values is provided, unless otherwise indicated, each intervening value to the tenth of the unit of the lower limit between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range is encompassed herein. The upper and lower limits of these smaller ranges may independently be included or excluded in the range, and each range where either, neither or both limits are included in the smaller ranges is also encompassed herein, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included.

[0046] The term “about” or “approximately” can mean within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation, per the practice in the given value. Where particular values are described in the application and claims, unless otherwise stated the term “about” should be assumed to mean an acceptable error range for the particular value, such as ±10% of the value modified by the term “about”.

[0047] The terms “individual,” “patient,” or “subject” can be used interchangeably. None of the terms require or are limited to situation characterized by the supervision (e.g. constant or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a physician’s assistant, an orderly, or a hospice worker). In some embodiments, patients, subjects, or individuals can be under the supervision of a health care worker.

[0048] The terms “heterologous nucleic acid sequence,” or “exogenous nucleic acid sequence,” or “transgenes,” as used herein, in relation to a specific virus can refer to a nucleic acid sequence that originates from a source other than the specified virus.

[0049] The terms “inhibiting,” “reducing” or “prevention,” or any variation of these terms, referred to herein, can include any measurable decrease or complete inhibition to achieve a desired result.

[0050] A “promoter,” as used herein, can be a control sequence that is a region of a nucleic acid sequence at which initiation and rate of transcription are controlled. In certain embodiments, a promoter may contain genetic elements at which regulatory proteins and molecules may bind such as RNA polymerase and other transcription factors. The terms “operatively positioned,” “operatively linked,” “under control” and “under transcriptional control” can mean that a promoter is in a correct functional location and / or orientation in relation to a nucleic acid sequence to control transcriptional initiation and / or expression of that sequence. In certain embodiments, a promoter may or may not be used in conjunction with an “enhancer,” which refers to a cis-acting regulatory sequence involved in the transcriptional activation of a nucleic acid sequence.

[0051] The term “percentage identical to,” as used herein, may be to calculations of “homology” or “percent homology” between two or more nucleotide or amino acid sequences that can be determined by aligning the sequences for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first sequence). The nucleotides at corresponding positions may then be compared, and the percent identity between the two sequences may be a function of the number of identical positions shared by the sequences (z.e., % homology = # of identical positions / total # of positions x 100). For example, a position in the first sequence may be occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent homology between the two sequences may be a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. In some embodiments, the length of a sequence aligned for comparison purposes may be at least about: 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 95%, of the length of the reference sequence. A BLAST® search may determine homology between two sequences. The homology can be between the entire lengths of two sequences or between fractions of the entire lengths of two sequences. The two sequences can be genes, nucleotides sequences, protein sequences, peptide sequences, amino acid sequences, or fragments thereof. The actual comparison of the two sequences can be accomplished by well-known methods, for example, using a mathematical algorithm. A non-limiting example of such a mathematical algorithm may be described in Karlin, S. and Altschul, S., Proc. Natl. Acad. Sci. USA, 90- 5873-5877 (1993). Such an algorithm may be incorporated into the NBLAST and XBLAST programs (version 2.0), as described in Altschul, S. et al., Nucleic Acids Res., 25:3389-3402 (1997). When utilizing BLAST and Gapped BLAST programs, any relevant parameters of the respective programs (e.g., NBLAST) can be used. For example, parameters for sequence comparison can be set at score= 100, word length= 12, or can be varied (e.g., W=5 or W=20). Other examples include the algorithm of Myers and Miller, CABIOS (1989), ADVANCE, ADAM, BLAT, and FASTA. In another embodiment, the percent identity between two amino acid sequences can be accomplished using, for example, the GAP program in the GCG software package (Accelrys, Cambridge, UK).

[0052] The term “subject” can refer to an animal, including, but not limited to, a primate (e.g., human), cow, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. The terms “subject” and “patient” are used interchangeably herein in reference, for example, to a mammalian subject, such as a human subject, or to a non-human subject.

[0053] The terms “treat,” “treating,” and “treatment” can be meant to include alleviating or abrogating a disorder, disease, or condition; or one or more of the symptoms associated with the disorder, disease, or condition; or alleviating or eradicating the cause(s) of the disorder, disease, or condition itself. Desirable effects of treatment can include, but are not limited to, preventing occurrence or recurrence of disease, alleviation of symptoms, diminishing any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state and remission or improved prognosis.

[0054] The term “therapeutically effective amount” can refer to the amount of a compound that, when administered, can be sufficient to prevent development of, or alleviate to some extent, one or more of the symptoms of the disorder, disease, or condition being treated. The term “therapeutically effective amount” can also refer to the amount of a compound that is sufficient to elicit the biological or medical response of a cell, tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor, or clinician.

[0055] The term “pharmaceutically acceptable carrier,” “pharmaceutically acceptable excipient,”“physiologically acceptable carrier,” or “physiologically acceptable excipient” can refer to a pharmaceutically-acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. A component can be “pharmaceutically acceptable” in the sense of being compatible with the other ingredients of a pharmaceutical formulation. It can also be suitable for use in contact with the tissue or organ of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications, commensurate with a reasonable benefit / risk ratio. ee, Remington: The Science and Practice of Pharmacy, 21st Edition; Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 5th Edition; Rowe et al., Eds., The Pharmaceutical Press and the American Pharmaceutical Association: 2005; and Handbook of Pharmaceutical Additives, 3rd Edition; Ash and Ash Eds., Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, Gibson Ed., CRC Press LLC: Boca Raton, FL, 2004).

[0056] The term “pharmaceutical composition” can refer to a mixture of a compound disclosed herein with other chemical components, such as diluents or carriers. The pharmaceutical composition can facilitate administration of the compound to an organism. Multiple techniques of administering a compound exist in the art including, but not limited to, oral, injection, aerosol, parenteral, and topical administration. Pharmaceutical compositions can also be obtained by reacting compounds with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p- toluenesulfonic acid, salicylic acid and the like.

[0057] The term “zika virus-like particle” can refer to a nanoscale multiprotein structure that is made up of assembled zika viral proteins and mimic the organization and conformation of the zika virus but without viral genetic material and are therefore non-infectious. The zika virus-like particle could closely resemble zika virus.

[0058] In some embodiments, as used here a “derivative” of a polypeptide or polynucleotide refers to a sequence at least 80% identical to the polypeptide or polynucleotide, respectively. In some embodiments, as used here a “derivative” of a polypeptide or polynucleotide refers to a sequence at least 80% homologous to the polypeptide or polynucleotide, respectively. In some embodiments, as used herein, a “derivative” of a polypeptide or polynucleotide refers to a sequence with no or no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid or nucleotide substitutions, insertions, or deletions as compared to the polypeptide or polynucleotide, respectively.Nucleic Acid Compositions

[0059] In one aspect, provided herein is a nucleic acid composition comprising (i) a polynucleotide encoding a zika virus capsid protein (C) or the derivative of the zika virus C, a polynucleotide encoding a zika virus viral membrane protein (prM / M) or the derivative of the zika virus prM / M, a polynucleotide encoding a zika virus viral envelope protein (E) or the derivative of the zika virus E, or any combination or two or more thereof; (ii) a polynucleotide encoding a zika virus C terminal viral envelope protein (E) or the derivative of the zika virus E, a polynucleotide encoding a zika virus non- structural protein 1 (NS1) or the derivative of the zika virus NS1, a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, a polynucleotide encoding a zika virus non- structural protein 2B (NS2B) or the derivative of the zika virus NS2B, a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3, a polynucleotide encoding a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, or any combination of two or more thereof; and (iii) a polynucleotide of interest. Also provided are nucleic acid compositions comprising (i) and (ii). Also provided are nucleic acid compositions comprising (i) and (iii). Also provided are nucleic acid compositions comprising (ii) and (iii). In some embodiments, (i) and (ii) are present on separate nucleic acids. In some embodiments, (i) and (ii) are present on the same nucleic acid.

[0060] In some embodiments, (i) is a first polynucleotide or a structural protein plasmid. In some embodiments, (ii) is a second polynucleotide or a non-structural protein plasmid. In some embodiments, (iii) is a third polynucleotide or a polynucleotide of interest plasmid. In some embodiments, (ii) is a helper plasmid that is not encapsulated by a virus-like particle. In some embodiments, (ii) is a polynucleotide that improves efficiency of virus-like particle assembly. In some embodiments, the nucleic acid composition comprises the derivative of the zika virus capsid protein (C), where the derivative encodes at least or about 5, 10, 15, 20, 25, or 30 amino acids of C. In some embodiments, the nucleic acid composition comprises the derivative of the zika virus C, where the derivative encodes about 5, 10, 15, 20, or 25 to about 30 amino acids of C. In some embodiments, the nucleic acid composition comprises a polynucleotide encoding complete C protein.

[0061] In one aspect, provided herein is a nucleic acid composition comprising a polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the polynucleotide does not encode one or more viral non-structural proteins and one or more viral structural proteins. In some embodiments, the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, and zika virus NS5 or a derivative of the zika virus NS5; and wherein the one or more viral structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, and zika virus viral envelope protein (E) or a derivative of the zika virus E. In some embodiments, each of the second polynucleotide and the third polynucleotide independently encode (i) a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus, and (ii) a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus. The polynucleotide may comprise a sequence encoding a protein of interest.

[0062] In some embodiments, the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, or zika virus NS5 or a derivative of the zika virus NS5, or a combination of two or more thereof. In some embodiments, the polynucleotide encodes a zika virus NS4a comprising a pTMS3. In some embodiments, the polynucleotide does not encode a zika virus NS4a comprising a pTMSl and a pTMS2. In some embodiments, the polynucleotide does not encode a full length zika virus NS4a. In some embodiments, the full length zika virus NS4a comprises a sequence at least 90% identical to SEQ ID NO: 10 or 2942. In some embodiments, the polynucleotide encodes a NS4b comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948. In some embodiments, the polynucleotide encodes a NS4b comprising a N-terminal disordered region. In some embodiments, the polynucleotide does not encode a zika virus NS4b comprising a pTMSl, a pTMS2, a pTMS3, a pTMS4, and a pTMS5. In some embodiments, the polynucleotide does not encode a full length zika virus NS4b. In some embodiments, the full length zika virus NS4b comprises a sequence at least 90% identical to SEQ ID NO: 12 or 2944. In some embodiments, the polynucleotide encodes a zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947.

[0063] In some embodiments, the one or more structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, zika virus viral envelope protein (E) or a derivative of the zika virus E, or a combination thereof. In some embodiments,the polynucleotide encodes a zika virus capsid protein (C) comprising a sequence at least 90% identical to any one of SEQ ID NOS: 2908-2917. In some embodiments, the polynucleotide does not encode a full length zika virus C. In some embodiments, the full length zika virus C comprises a sequence at least 90% identical to SEQ ID NO: 3 or 2901. In some embodiments, the zika virus-like particle comprises the one or more viral non- structural proteins and the one or more viral structural proteins, optionally wherein the one or more structural proteins are assembled into a zika virus-like shell, and further optionally wherein the zika virus-like particle is not replication competent.

[0064] In one aspect, provided herein is a composition comprising: (a) a first polynucleotide encoding a zika virus structural protein, (b) a second polynucleotide encoding a zika virus non- structural (NS) protein, and (c) a third polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the first polynucleotide and the second polynucleotide independently do not encode the zika virus 5’ UTR or the zika virus 3’ UTR. The third polynucleotide may comprise a sequence encoding a protein of interest.

[0065] In some embodiments, the first polynucleotide encoding the zika virus structural protein comprises: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C; (d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; (e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E, or (f) a combination of two or more selected from (a) to (e). In some embodiments, the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919. In some embodiments, the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0066] In some embodiments, the second polynucleotide encoding a zika virus non- structural (NS) protein comprises: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1; (d) a polynucleotide encoding a zika virus non- structural protein 2 A (NS2A) or the derivative of the zika virus NS2A; (e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A;(f) a polynucleotide encoding a zika virus non- structural protein 2B (NS2B) or the derivative of the zika virus NS2B; (g) a polynucleotide encoding a zika virus non- structural protein 3 (NS3) or the derivative of the zika virus NS3; (h) a polynucleotide encoding a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A; (i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K; (j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B; (k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5; or (1) a combination of two or more selected from (a) to (k). In some embodiments, the second polypeptide further comprises a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E. In some embodiments, the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919. In some embodiments, the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0067] In some embodiments, the third polynucleotide encoding a zika virus 5’ UTR of Table IB and a zika virus 3’ UTR of Table IB comprises: (a) a polynucleotide encoding a zika virus 5’ UTR of the zika nucleotide sequences of Table IB; (b) a polynucleotide encoding a zika virus 3’ UTR of the zika nucleotide sequences of Table IB; (c) a polynucleotide of interest; (d) a non- structural protein, or a derivative of a non-structural protein; (e) a structural protein, or a derivative of a structural protein; or (f) a combination of two or more selected from (a) to (e). In some embodiments, the non-structural protein comprises a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A, a polynucleotide encoding a zika virus non- structural protein 2K (2K) or the derivative of the zika virus 2K, and a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B. In some embodiments, the non-structural protein comprises a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A comprising a sequence at least 90% identical to SEQ ID NO: 2929 or 2946, a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947, and a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948. In some embodiments, the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C. In some embodiments, the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C comprising a sequence at least 90% identical to SEQ ID NOS: 2908- 2917.

[0068] In one aspect, provided herein is a structural protein plasmid comprising: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C; (d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; and (e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E; wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919, and wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0069] In one aspect, provided herein is a non-structural protein plasmid comprising: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1; (d) a polynucleotide encoding a zika virus non-structural protein 2A (NS2A) or the derivative of the zika virus NS2A; (e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A; (f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B; (g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3; (h) a polynucleotide encoding a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A; (i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K; (j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B; (k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5; (1) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E; or (m) a combination of two or more selected from (a) to (1); wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919, and wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0070] In one aspect, provided herein is a polynucleotide of interest plasmid comprising: (a) a polynucleotide encoding a zika virus 5’ UTR of the zika nucleotide sequences of Table IB; (b) a polynucleotide encoding a zika virus 3’ UTR of the zika nucleotide sequences of Table IB; (c) apolynucleotide of interest; (d) a non-structural protein, or a derivative of a non-structural protein; (e) a structural protein, or a derivative of a structural protein; or (f) a combination of two or more selected from (a) to (e); wherein the non-structural protein comprises a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, and a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, and wherein the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C.Structural and non-structural proteins from zika virus

[0071] A wild-type zika virus genome comprises a 10.8-kilobase single-stranded positive-sense RNA that codes for three structural proteins (C, prM / M, and envelope E) and seven non- structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5). In addition, there are short UTRs on both the 5' and 3' ends of the genome. Provided herein are various embodiments to generate a zika virus-like particle that comprises at least one or all of the three structural proteins. In some embodiments, a nucleic acid composition comprising one or more polynucleotides is provided for generating the zika virus-like particle, wherein at least one of the polynucleotides encodes a polynucleotide of interest for delivery by the zika virus-like particle.

[0072] In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., a structural protein plasmid) comprises the polynucleotide encoding the zika virus C, the polynucleotide encoding the zika virus prM / M, and the polynucleotide encoding the zika virus E. When the three structural proteins are expressed as the nucleic acid composition described herein, in some embodiments, they could be expressed in one nucleic acid. When three structural proteins are expressed as the nucleic acid composition described herein, in other embodiments, they could be expressed in more than one nucleic acids.

[0073] In one aspect, provided herein is a nucleic acid composition comprising a polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the polynucleotide does not encode one or more viral non-structural proteins and one or more viral structural proteins. In some embodiments, the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, and zika virus NS5 or a derivative of the zika virus NS5; and wherein the one or more viral structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, and zika virus viral envelope protein (E) or a derivative of the zika virus E. In some embodiments, each of the second polynucleotide and the third polynucleotide independently encode (i) a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90%identical to the 5’ UTR of the zika virus, and (ii) a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus.

[0074] In some embodiments, the one or more viral non- structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, or zika virus NS5 or a derivative of the zika virus NS5, or a combination of two or more thereof. In some embodiments, the polynucleotide encodes a zika virus NS4a comprising a pTMS3. In some embodiments, the polynucleotide does not encode a zika virus NS4a comprising a pTMSl and a pTMS2. In some embodiments, the polynucleotide does not encode a full length zika virus NS4a. In some embodiments, the full length zika virus NS4a comprises a sequence at least 90% identical to SEQ ID NO: 10 or 2942. In some embodiments, the polynucleotide encodes a NS4b comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948. In some embodiments, the polynucleotide encodes a NS4b comprising a N-terminal disordered region. In some embodiments, the polynucleotide does not encode a zika virus NS4b comprising a pTMSl, a pTMS2, a pTMS3, a pTMS4, and a pTMS5. In some embodiments, the polynucleotide does not encode a full length zika virus NS4b. In some embodiments, the full length zika virus NS4b comprises a sequence at least 90% identical to SEQ ID NO: 12 or 2944. In some embodiments, the polynucleotide encodes a zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947.

[0075] In some embodiments, the one or more structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, zika virus viral envelope protein (E) or a derivative of the zika virus E, or a combination thereof. In some embodiments, the polynucleotide encodes a zika virus capsid protein (C) comprising a sequence at least 90% identical to any one of SEQ ID NOS: 2908-2917. In some embodiments, the polynucleotide does not encode a full length zika virus C. In some embodiments, the full length zika virus C comprises a sequence at least 90% identical to SEQ ID NO: 3 or 2901. In some embodiments, the zika virus-like particle comprises the one or more viral non- structural proteins and the one or more viral structural proteins, optionally wherein the one or more structural proteins are assembled into a zika virus-like shell, and further optionally wherein the zika virus-like particle is not replication competent.

[0076] In some embodiments, the first polynucleotide encoding the zika virus structural protein comprises: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR ofthe zika virus; (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C; (d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; (e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E, or (f) a combination of two or more selected from (a) to (e). In some embodiments, the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919. In some embodiments, the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0077] In some embodiments, the second polynucleotide encoding a zika virus non- structural (NS) protein comprises: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1; (d) a polynucleotide encoding a zika virus non- structural protein 2 A (NS2A) or the derivative of the zika virus NS2A; (e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A; (f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B; (g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3; (h) a polynucleotide encoding a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A; (i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K; (j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B; (k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5; or (1) a combination of two or more selected from (a) to (k). In some embodiments, the second polypeptide further comprises a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E. In some embodiments, the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919. In some embodiments, the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0078] In one aspect, provided herein is a structural protein plasmid comprising: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C; (d) apolynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; and (e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E; wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919, and wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0079] In one aspect, provided herein is a non-structural protein plasmid comprising: (a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus; (b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus; (c) a polynucleotide encoding a zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1; (d) a polynucleotide encoding a zika virus non-structural protein 2A (NS2A) or the derivative of the zika virus NS2A; (e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A; (f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B; (g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3; (h) a polynucleotide encoding a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A; (i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K; (j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B; (k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5; (1) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E; or (m) a combination of two or more selected from (a) to (1); wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919, and wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

[0080] In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) encodes a zika virus C or a derivative of the zika virus C.

[0081] In some embodiments, the polynucleotide encoding the derivative of the zika virus C comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus C. In specific embodiments, the zika virus C or the derivative of the zika virus C comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99%, or 100% identical to a sequence of Table 1A, Table 2, or Table 3 or a zika virus C of a zika virus from Table IB. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus C encoding an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a wild type C sequence. In some embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus C encoding an amino acid sequence at least 95% identical to a wild type C sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus C encoding an amino acid sequence at least 96% identical to a wild type C sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus C encoding an amino acid sequence at least 97% identical to a wild type C sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus C encoding an amino acid sequence at least 98% identical to a wild type C sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus C encoding an amino acid sequence at least 99% identical to a wild type C sequence. In some embodiments, the nucleic acid composition or polynucleotide comprises a derivative of zika virus C encoding about 5 to about 30 amino acids of a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence of Table 1A, Table 2, Table 3, or a wild-type zika virus of Table IB.

[0082] In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) encodes a zika virus prM / M or a derivative of the zika virus prM / M. In some embodiments, the polynucleotide encoding the derivative of the zika virus prM / M comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus prM / M. In some embodiments, the zika virus prM / M or the derivative of the zika virus prM / M comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence of Table 1A or a prM / M of a zika virus from Table IB. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus prM / M encoding an amino acid sequence at least 95% identical to a wild type prM / M sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus prM / M encoding an amino acid sequence at least 96% identical to a wild type prM / M sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus prM / M encoding an amino acid sequence at least 97%identical to a wild type prM / M sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus prM / M encoding an amino acid sequence at least 98% identical to a wild type prM / M sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus prM / M encoding an amino acid sequence at least 99% identical to a wild type prM / M sequence.

[0083] In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) encodes a zika virus E or a derivative of the zika virus E. In some embodiments, the polynucleotide encoding the derivative of the zika virus E comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus E. In some embodiments, the polynucleotide encoding E is translated into a wild zika virus type E. In some embodiments, the zika virus E or the derivative of the zika virus E comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 1A, or an E of a zika virus from Table IB. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika E encoding an amino acid sequence at least 95% identical to a wild type E sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus E encoding an amino acid sequence at least 96% identical to a wild type E sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus E encoding an amino acid sequence at least 97% identical to a wild type E sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus E encoding an amino acid sequence at least 98% identical to a wild type E sequence. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus E encoding an amino acid sequence at least 99% identical to a wild type E sequence.

[0084] In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) comprises a 5’ untranslated region (5’ UTR). In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non- structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) comprises a 3’ untranslated region (3’ UTR). Non-limiting example untranslated sequences are provided in Table 1 A, Table IB, and Table 2, and the UTRs of zika viruses in Table IB. In some embodiments, the 5’ UTR comprises a sequence at least 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 1A or Table 2, or a 5’ UTR of a zika virus from Table IB. In some embodiments, the 3’ UTR comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 1A, Table IB, or Table 2, or a 3’ UTR of a zika virus from Table IB. In some embodiments, a structural protein plasmid or first polynucleotide comprises a 5’ UTR and a 3’ UTR, where both 5’ and 3’ UTRs are not UTRs of a zika virus. In some embodiments, a non- structural protein plasmid or second polynucleotide comprises a 5’ UTR and a 3’ UTR, where both 5’ and 3’ UTRs are not UTRs of a zika virus. In some embodiments the 5’ UTR is a 5’ UTR exogenous to the zika virus. Non-limiting examples of exogenous 5’ UTRs are provided in Table 1C. In some embodiments, the 5’ UTR exogenous to the zika virus comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 1C. In some embodiments the 3’ UTR is a 3’ UTR exogenous to the zika virus. Non-limiting examples of exogenous 3’ UTRs are provided in Table ID. In some embodiments, the 3’ UTR exogenous to the zika virus comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table ID.

[0085] In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, polynucleotide of interest plasmid or third polynucleotide) does not comprise a polynucleotide encoding one or more non-structural (NS) proteins selected from (i) a NS1, (ii) a NS2A, (iii) a NS2B, (iv) a NS3, (v) a NS4A, (vi) a NS4B, (vii) a NS5, or (viii) two or more of (i)-(vii). For example, if the nucleic acid does not comprise such polynucleotide(s), there is additional space in the composition to accommodate a large polynucleotide of interest (e.g., up to about 10000 or 6000 bases).

[0086] In other embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises polynucleotide encoding one or more non-structural (NS) proteins selected from (i) a NS1, (ii) a NS2A, (iii) a NS2B, (iv) a NS3, (v) a NS4A, (vi) a NS4B, (vii) a NS5, or (viii) two or more of (i)-(vii). In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envel op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0087] In some embodiments, the zika virus NS4a or the derivative of the zika virus NS4a comprises an amino acid sequence at least 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence of Table 1A, Table IB, Table 4, Table 5, Table 8, or Table 9. In specific embodiments, the nucleic acid composition or polynucleotide comprises aderivative of the zika virus NS4a encoding an amino acid sequence at least 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a wild type NS4a sequence. In some embodiments, the zika virus NS4a or the derivative of the zika virus NS4a is a partial zika virus NS4a. In some embodiments, the partial NS4a comprises a pTMS3 (putative transmembrane sequence 3). In some embodiments, the partial NS4a does not comprise a pTMSl and a pTMS2. In some embodiments, the partial NS4a comprises a SEQ ID NO: 2929. In some embodiments, the partial NS4a comprises a SEQ ID NO: 2946.

[0088] In some embodiments, the zika virus NS4b or the derivative of the zika virus NS4b comprises an amino acid sequence at least 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence of Table 1A, Table IB, Table 4, Table 5, Table 8, or Table 9. In specific embodiments, the nucleic acid composition or polynucleotide comprises a derivative of the zika virus NS4b encoding an amino acid sequence at least 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a wild type NS4b sequence. In some embodiments, the zika virus NS4b or the derivative of the zika virus NS4b is a partial zika virus NS4b. In some embodiments, the partial NS4a comprises a N-terminal disordered region. In some embodiments, the partial NS4a does not comprise a pTMSl, a pTMS2, a pTMS3, a pTMS4, and a pTMS5. In some embodiments, the partial NS4a comprises a SEQ ID NO: 2931. In some embodiments, the partial NS4a comprises a SEQ ID NO: 2948.

[0089] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1 protein. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A protein. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B protein. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS3 protein. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS4A protein. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS4B protein. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS5 protein. In certain embodiments, thenucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envel op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0090] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1 and a NS2A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1 and a NS2B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1 and a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1 and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1 and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1 and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0091] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A and a NS2B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A and a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certainembodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0092] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B and a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0093] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS3 and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS3 and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS3 and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0094] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS4A and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS4A and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0095] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS4B and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0096] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2A, and NS2B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2A, and NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2A, and NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2A, and NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2A, and NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0097] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2B, and NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2B, and NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2B, and NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS2B, and NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0098] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS3, and NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS3, and NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS3, and NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (env el op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0099] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS4A, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NSl, a NS4A, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envel op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0100] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS1, a NS4B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0101] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS2B, and a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS2B, and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS2B, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotideencoding a NS2A, a NS2B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0102] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS3, and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS3, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS3, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0103] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS4A, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS4A, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0104] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2A, a NS4B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0105] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B, a NS3, and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide)comprises a polynucleotide encoding a NS2B, a NS3, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B, a NS3, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envel op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0106] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B, a NS4A, and NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0107] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS2B, a NS4B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0108] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS3 and a NS4A, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0109] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS3, a NS4B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0110] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a NS4A, a NS4B, and a NS5. In certain embodiments, the nucleic acid compositioncomprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0111] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS2A, and NS2B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS2A, and NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS2A, and NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NSl, a NS2A, and NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NSl, a NS2A, and NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0112] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS2B, and NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS2B, and NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS2B, and NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS2B, and NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0113] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS3, and NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS3, and NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS3, and NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (env el op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0114] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS4A, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS4A, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0115] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1, a NS4B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0116] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A, a NS2B, and a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A, a NS2B, and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide)comprises a polynucleotide encoding all NS proteins except a NS2A, a NS2B, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide). In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envel op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0117] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A, a NS3, and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A, a NS3, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A, a NS3, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envel op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0118] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A, a NS4A, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A, a NS4A, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0119] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A, a NS4B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0120] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B, a NS3, and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B, a NS3, and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B, a NS3, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (env el op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0121] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B, a NS4A, and NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0122] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except all NS proteins except a NS2B, a NS4B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0123] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS3 and a NS4A, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0124] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS3, a NS4B, and a NS5. In certain embodiments, the nucleicacid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0125] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS4A, a NS4B, and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0126] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1 and a NS2A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NSl and a NS2B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1 and a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except all NS proteins except a NS1 and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1 and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NSl and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0127] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A and a NS2B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A and a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g.,non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0128] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B and a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0129] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS3 and a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS3 and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS3 and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments thederivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0130] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS4A and a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS4A and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0131] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS4B and a NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0132] In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS1. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS2B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS3. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS4A. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS4B. In certain embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a polynucleotide encoding all NS proteins except a NS5. In certain embodiments, the nucleic acid compositioncomprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0133] In some embodiments, the NS1 is a zika virus NS1. In some embodiments, the NS2A is a zika virus NS2A. In some embodiments, the NS2B is a zika virus NS2B. In some embodiments, the NS3 is a zika virus NS3. In some embodiments, the NS4A is a zika virus NS4A. In some embodiments, the NS4B is a zika virus NS4B. In some embodiments, the NS5 is a zika virus NS5. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (envelop-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0134] In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a derivative of NS1 that comprises an amino acid sequence at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a wild-type NS1 sequence of Table 1 A or from a zika virus of Table IB. In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a derivative of NS2A that comprises an amino acid sequence at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a wild-type NS2A sequence of Table 1A or from a zika virus of Table IB. In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a derivative of NS2B that comprises an amino acid sequence at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a wild-type NS2B sequence of Table 1 A or from a zika virus of Table IB. In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a derivative of NS3 that comprises an amino acid sequence at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a wild-type NS3 sequence of Table 1A or from a zika virus of Table IB. In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non- structural protein plasmid or second polynucleotide) comprises a derivative of NS4A that comprises an amino acid sequence at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a wild-type NS4A sequence of Table 1A or from a zika virus of Table IB. In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises aderivative of NS4B that comprises an amino acid sequence at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a wild-type NS4B sequence of Table 1A or from a zika virus of Table IB. In some embodiments, the nucleic acid composition or a polynucleotide described herein (e.g., non-structural protein plasmid or second polynucleotide) comprises a derivative of NS5 that comprises an amino acid sequence at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a wild-type NS5 sequence of Table 1 A or from a zika virus of Table IB. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding a E protein or a derivative of E. In certain embodiments the derivative of E (env el op-partial) comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table 3.

[0135] In some embodiments, the components of the nucleic acid composition comprise African zika virus components, Asian zika virus components, or Brazilian zika virus components, or a combination thereof. In one specific embodiment, the zika virus is African MR766 strain.Regulatory regions of zika genome

[0136] A wild type zika genome comprises several regulatory regions, such as 5’ UTR and 3’ UTR. UTRs are found to be important in viral replication and immune modulation. Elements in UTRs are found to be essential for genome cyclization, resulting in initiation of RNA synthesis.

[0137] Ribozymes are self-cleaving RNAs. Small ribozyme motifs mainly fall within four types: hammerhead, hairpin, Varkud satellite (VS), and hepatitis delta virus (HDV).

[0138] In some embodiments, a nucleic acid composition or a polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) comprises a polynucleotide encoding a ribozyme.

[0139] In one aspect, provided herein is a nucleic acid composition or a polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) comprises a polynucleotide encoding a ribozyme. In some embodiments, the ribozyme is a hammerhead ribozyme or a HDV ribozyme. In some embodiments, the nucleic acid or polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) further comprises a polynucleotide encoding a second ribozyme. In some specific embodiments, the second ribozyme is a hammerhead ribozyme or a hepatitis delta virus (HDV) ribozyme. Insome embodiments, the polynucleotide encoding the ribozyme encodes a hammerhead ribozyme and the polynucleotide encoding the second ribozyme encodes an HDV ribozyme.Additional features of a nucleic acid composition

[0140] In some embodiments, the nucleic acid composition or polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) further comprises one or more expression control elements in operable linkage that confers expression of the nucleic acid composition in vitro. In some embodiments, the expression control element is a promoter that drives expression of the nucleic acid complex in vitro. In specific embodiments, the promoter is T7, T3, SP6 or any phage promoter.

[0141] In another aspect, provided herein is a pharmaceutical composition comprising the nucleic acid composition or polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non-structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) described herein with a pharmaceutically acceptable salt or derivative thereof.Polynucleotides of Interest

[0142] In some embodiments, a polynucleotide herein, e.g., of a plasmid such as a pREP plasmid, comprises a polynucleotide of interest. In some embodiments, a protein of interest is encoded by a polynucleotide of interest. In some embodiments, the polynucleotide of interest is about or up to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides long. For example, the polynucleotide of interest is about 50 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides long. The polynucleotide of interest may be about 100 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides long. The polynucleotide of interest may be about 50 to about 6000 nucleotides long. The polynucleotide of interest may be about 100 to about 6000 nucleotides long. The polynucleotide of interest may be about 50 to about 5000 nucleotides long. The polynucleotide of interest may be about 100 to about 5000 nucleotides long. In some embodiments, the polynucleotide of interest is selected from Table IE. In some embodiments, the polynucleotide of interest comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table IE.

[0143] In some embodiments, the polynucleotide of interest is about 0.1 kb nucleotides long. In other embodiments, the polynucleotide of interest is about 0.2 kb long. In other embodiments, the polynucleotide of interest is about 0.3 kb long. In other embodiments, the polynucleotide of interest is about 0.4 kb long. In other embodiments, the polynucleotide of interest is about 0.5 kb long. In other embodiments, the polynucleotide of interest is about 0.6 kb long. In otherembodiments, the polynucleotide of interest is about 0.7 kb long. In other embodiments, the polynucleotide of interest is about 0.8 kb long. In other embodiments, the polynucleotide of interest is about 0.9 kb long. In other embodiments, the polynucleotide of interest is about 1 kb nucleotides long. In other embodiments, the polynucleotide of interest is about 2 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 3 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 4 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 5 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 6 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 7 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 8 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 9 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 10 kb nucleotides long. In some embodiments, the polynucleotide of interest is selected from Table IE. In some embodiments, the polynucleotide of interest comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table IE.

[0144] In some embodiments, the polynucleotide of interest is an antigen or an antigenic epitope thereof. In some embodiments, the antigen or an antigenic epitope thereof is disease associated. In specific embodiments, the antigen or an antigenic epitope thereof is from a pathogen.

[0145] In some embodiments, the pathogen is a virus. In specific embodiments, the virus is a human SARS coronavirus, an influenza A virus, an influenza B virus, an influenza C virus an ebolavirus, a hepatitis B virus, a hepatitis C virus, a herpes simplex virus, a human immunodeficiency virus (HIV), a human papillomavirus (HPV-6, HPV-11), a measles virus, a rabies virus, a poliovirus, or a yellow fever virus.

[0146] In some embodiments, the pathogen is a bacterium. In specific embodiments, the bacteria are Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella. Henselae, Bartonella quintana, Bifidobacterium Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, Cardiobacterium hominis, Campylobacter fetus, Chlamydia pneumonia, Chlymydia trahomatis, Clostridium difficile, Cyanobacteria, Eikennella corrodens, Enterobacter, Enterococcus faccium, Escherichia coli, Escherichia coli 0157, Franceilla tularensis, Fusobacterium nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzae, Helicobacter pylori, Kingella kingae, Klebsiella pneumonia, Legionella bacteria, Legionella pneumophila serogroup 1, Leptospria, Morganella morganii, Neisseria gonorrhoeae, Neisseria meningitidis, Proteus mirabilis, Proteus vulgaris, Proteus myxofaciens, Providenciarettgeri, Providencia alcalifaciens, Providencia stuartii, Pseudomonas aeruginosa, Pseudomonas paucimobilis, Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas acidovorans, Rickettsiae, Salmonella enterica, Salmonella typhi, Salmonella paratyphi types A, B typhus, Salmonella, Salmonella arizonae, Salmonella choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonnei, Treponema, Stenotrophomonas maltophilia, Vibrio cholerae, Vibrio mimicus, Vibrio alginolyticus, Vibrio hollisae, Vibrio parahaemolyticus, Vibrio vulnificus, Yersinia pestitis, Actinomycetes, Bacillus anthracis, Bacillus subtilis, Clostridium tetani, Clostridium. Perfingens, Clostridium botulinum, Clostridium tetani. Corynebacterium diphtheriae, Enterococcus faecalis, Enterococcus faecium, Erysipelothrix ruhsiopathiae, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium tuberculosis, Mycoplasma, Nocardia, Propionibacerium, Pseudomonas aeruginosa, Pneumococci, Staphylococcus aureus, Staphylococcus epidermidis, methicillin resistant Staphylococcus aureus (MRSA), vancomycin resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcus saprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutants.

[0147] In some embodiments, the pathogen is a fungus, an amoeba, or a parasite. In specific embodiments, the fungus, the amoeba, or the parasite is Acanthamoeba spp, American tryppanosomiasis, Balamuthia mandnillanis, Babesia divergenes, Babesia bigemina, Babesia equi, Babesia microfti, Babesia duncani, Balantidium coli, Blastocystis spp Cryptosporidium spp, Cyclospora cayetanensis, dientamoeba fragilis, Diphyllobothrium latum, Leishmania amazonesis, Naegleria fowderi, Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale curtisi, Plasmodium malariae, Rhinosporidium seeberi, Sarcocystis bovihominis, Sarcocystiss suihominis, Toxoplasma gondii, Trichmonas vaginalis, Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0148] In some embodiments, the polynucleotide of interest encodes a gene editing tool. In specific embodiments, the gene editing tool is selected from a group comprising meganuclease associated agents, CRISPR associated agents, TALEN associated agents, and zinc finger associated agents.

[0149] In some embodiments, the polynucleotide of interest is a small interfering RNA (siRNA), antisense RNA, micro RNA (miRNA), small or short hairpin RNA (shRNA), guide RNA (gRNA), clustered regularly interspaced short palindromic repeat RNA (crRNA), trans-activating clustered regularly interspaced short palindromic repeat RNA (tracrRNA), immune-stimulating oligonucleotide, antisense nucleic acid, or ribozyme. In specific embodiments, the polynucleotide of interest targets beta-secretase 1 (BACE1) and / or amyloid precursor protein (APP).

[0150] In some embodiments, the polynucleotide of interest encodes a polypeptide associated with a genetic disorder. In specific embodiments, the polypeptide is brain-derived neurotrophin factor (BDNF), nerve growth factor (NGF), Neprilysin inhibitors (NEP), endothelin converting enzyme (ECE), cathepsin B (CTSB), apolipoprotein E 2 (APOE2), SH3 and multiple ankyrin repeat domains protein (SHANK), neurturin (NRTN), glial cell-derived neurotrophic factor (GDNF), cerebral dopamine neurotrophic factor (CDNF), vascular endothelial growth factor A (VEGF-A), or aromatic L-amino acid decarboxylase (AADC).

[0151] In some embodiments, the polynucleotide of interest encodes a therapeutic agent or a diagnostic agent. In specific embodiments, the therapeutic agent is an antibody-based therapeutic agent, a hormone, a cytokine, an inhibitor or antagonist of an immune checkpoint regulator, an immune stimulatory molecule, or an agonist of an immune co-stimulatory molecule. In further embodiments, the inhibitor or antagonist of an immune checkpoint regulator is an anti-PDl antibody. In another embodiments, the antibody -based therapeutic agent is an antibody, a functional fragment of an antibody, a chimeric antigen receptor (CAR), or a T cell receptor (TCR). In still other embodiments, the cytokine comprises lymphokine, monokine, polypeptide hormone, growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, glycoprotein hormone, follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), hepatic growth factor, fibroblast growth factor, prolactin, placental lactogen, tumor necrosis factor-a and -P, inhibiting substance, mouse gonadotropin-associated peptide, inhibin, activin, vascular endothelial growth factor, integrin, thrombopoietin (TPO), nerve growth factor, NGF-P, platelet growth factor, transforming growth factor (TGF), TGF-a, TGF-P, insulin-like growth factor-1 and -II, erythropoietin (EPO), osteoinductive factor, interferon, such as interferon-a, -P and -y, colony stimulating factor (CSF), macrophage-CSF (M-CSF), granulocyte-macrophage-CSF (GM-CSF), granulocyte-CSF (GCSF), interleukin (IL), tumor necrosis factor, TNF-a, TNF-P, LIF, or kit ligand (KL), or a combination of two or more thereof.

[0152] In some embodiments, the polynucleotide of interest is a reporter. In some embodiments the polynucleotide of interest encodes for NanoLuc, GFP, beta-galactosidase, or luciferase. In some embodiments, the polynucleotide of interest encodes for NanoLuc. In some embodiments, the polynucleotide of interest encodes for GFP. In some embodiments, the polynucleotide of interest encodes for beta-galactosidase. In some embodiments, the polynucleotide of interest encodes for luciferase. In some embodiments, the polynucleotide of interest encodes for NanoLuc, GFP, beta-galactosidase, luciferase, or a combination of two or more thereof. In some embodiments the polynucleotide of interest is selected from Table IE. In some embodiments, thepolynucleotide of interest comprises a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence of Table IE.

[0153] In some embodiments, the polynucleotide of interest is about or up to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides long. For example, the polynucleotide of interest is about 50 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides long. The polynucleotide of interest may be about 100 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides long. The polynucleotide of interest may be about 50 to about 6000 nucleotides long. The polynucleotide of interest may be about 100 to about 6000 nucleotides long. The polynucleotide of interest may be about 50 to about 5000 nucleotides long. The polynucleotide of interest may be about 100 to about 5000 nucleotides long.

[0154] In some embodiments, the polynucleotide of interest is about 0.1 kb nucleotides long. In other embodiments, the polynucleotide of interest sequence is about 0.2 kb long. In other embodiments, the polynucleotide of interest sequence is about 0.3 kb long. In other embodiments, the polynucleotide of interest sequence is about 0.4 kb long. In other embodiments, the polynucleotide of interest sequence is about 0.5 kb long. In other embodiments, the polynucleotide of interest sequence is about 0.6 kb long. In other embodiments, the polynucleotide of interest sequence is about 0.7 kb long. In other embodiments, the polynucleotide of interest sequence is about 0.8 kb long. In other embodiments, the polynucleotide of interest sequence is about 0.9 kb long. In other embodiments, the polynucleotide of interest sequence is about 1 kb nucleotides long. In other embodiments, the polynucleotide of interest polynucleotide is about 2 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 3 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 4 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 5 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 6 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 7 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 8 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 9 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 10 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 11 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 12 kb nucleotides long. In some embodiments, polynucleotide of interest is about 13 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 14 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 15 kb nucleotides long. In some embodiments, thepolynucleotide of interest is about 16 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 17 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 18 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 19 kb nucleotides long. In some embodiments, the polynucleotide of interest is about 20 kb nucleotides long.Additional example features

[0155] In some embodiments, components of the nucleic acid composition are expressed on one nucleic acid, or two or more separate nucleic acids. In an example embodiment, the nucleic acid composition comprises a first polynucleotide, a second polynucleotide, and a third polynucleotide. In some embodiments, the first polynucleotide, second polynucleotide, and third polynucleotides are present on separate nucleic acids. In an example embodiment, the nucleic acid composition comprises a structural protein plasmid, a non-structural protein plasmid, and a polynucleotide of interest plasmid. In an example embodiment, the nucleic acid composition comprises a structural protein plasmid and a non-structural protein plasmid. In an example embodiment, the nucleic acid composition comprises a structural protein plasmid and a polynucleotide of interest plasmid. In an example embodiment, the nucleic acid composition comprises a non-structural protein plasmid, and a polynucleotide of interest plasmid. In some embodiments, the nucleic acid composition, first polynucleotide, second polynucleotide, and / or third polynucleotide further comprises one or more expression control elements in operable linkage that confers expression of the nucleic acid composition in vitro or in vivo. In some embodiments, the expression control element is a promoter that drives expression of the nucleic acid complex in vitro. In specific embodiments, the promoter is T7, T3, SP6 or any phage promoter.

[0156] In some embodiments, the expression control element is a promoter that drives expression of the nucleic acid complex in a target cell. In some specific embodiments, the promoter is CMV, SV40 or any eukaryotic promoter.

[0157] In some embodiments, the target cell is a neuron, or a non-neuron cell, VERO, COS, CHO, C6 / 36, HeLa, HEK, or HepG2. In some embodiments, the target cell is an oligodendrocyte, microglia, or astrocyte.

[0158] In some embodiments, provided herein is a pharmaceutical composition comprising the nucleic acid composition described above with a pharmaceutically acceptable salt or derivative thereof.Zika virus-like particles

[0159] Viral vectors are widely used in fields such as gene therapy. Currently, the three key vectors are based on adenoviruses, adeno-associated viruses, and lentiviruses. However, limitations for each vectors exist. Provided herein is a zika virus-like particle which in some embodiments provides more flexible packaging strategies, similar or better delivery efficiency, decreased insertional mutagenesis, decreased immune response, safety, or special tropism, or any combination thereof.

[0160] In some embodiments, the zika virus-like particle is generated from expressing the nucleic acid composition or polynucleotide described herein (e.g., structural protein plasmid or first polynucleotide, non- structural protein plasmid or second polynucleotide, polynucleotide of interest plasmid or third polynucleotide) in a producer cell.

[0161] In specific embodiments, the zika virus-like particle is produced from a nucleic acid composition comprising a structural protein plasmid or first polynucleotide, a non- structural protein plasmid or second polynucleotide, and a polynucleotide of interest plasmid or third polynucleotide. The structural protein plasmid or first polynucleotide may comprise a polynucleotide encoding a zika virus capsid protein (C) or the derivative of the zika virus C, a polynucleotide encoding a zika virus viral membrane protein (prM / M) or the derivative of the zika virus prM / M, a polynucleotide encoding a zika virus viral envelope protein (E) or the derivative of the zika virus E, or any combination or two or more thereof. The non- structural protein plasmid or second polynucleotide may comprise a polynucleotide encoding a zika virus C terminal viral envelope protein (E) or the derivative of the zika virus E, a polynucleotide encoding a zika virus non- structural protein 1 (NS 1) or the derivative of the zika virus NS1, a polynucleotide encoding a zika virus non- structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B, a polynucleotide encoding a zika virus non- structural protein 3 (NS3) or the derivative of the zika virus NS3, a polynucleotide encoding a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, or any combination of two or more thereof. The polynucleotide of interest plasmid or third polynucleotide may comprise a polynucleotide of interest.

[0162] In specific embodiments, the zika virus-like particle is produced from a nucleic acid composition comprising a structural protein plasmid or first polynucleotide and a non- structural protein plasmid or second polynucleotide. The structural protein plasmid or first polynucleotide may comprise a polynucleotide encoding a zika virus capsid protein (C) or the derivative of the zika virus C, a polynucleotide encoding a zika virus viral membrane protein (prM / M) or the derivative of the zika virus prM / M, a polynucleotide encoding a zika virus viral envelope protein (E) or the derivative of the zika virus E, or any combination or two or more thereof. The non- structural protein plasmid or second polynucleotide may comprise a polynucleotide encoding a zika virus C terminal viral envelope protein (E) or the derivative of the zika virus E, a polynucleotide encoding a zika virus non- structural protein 1 (NS1) or the derivative of the zika virus NS1, a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, a polynucleotide encoding a zika virus non- structural protein 2B (NS2B) or the derivative of the zika virus NS2B, a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3, a polynucleotide encoding a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, or any combination of two or more thereof.

[0163] In specific embodiments, the zika virus-like particle is produced from a nucleic acid composition comprising a structural protein plasmid or first polynucleotide and a polynucleotide of interest plasmid or third polynucleotide. The structural protein plasmid or first polynucleotide may comprise a polynucleotide encoding a zika virus capsid protein (C) or the derivative of the zika virus C, a polynucleotide encoding a zika virus viral membrane protein (prM / M) or the derivative of the zika virus prM / M, a polynucleotide encoding a zika virus viral envelope protein (E) or the derivative of the zika virus E, or any combination or two or more thereof. The polynucleotide of interest plasmid or third polynucleotide may comprise a polynucleotide of interest.

[0164] In specific embodiments, the zika virus-like particle is produced from a nucleic acid composition comprising a non-structural protein plasmid or second polynucleotide and a polynucleotide of interest plasmid or third polynucleotide. The non-structural protein plasmid or second polynucleotide may comprise a polynucleotide encoding a zika virus C terminal viralenvelope protein (E) or the derivative of the zika virus E, a polynucleotide encoding a zika virus non- structural protein 1 (NS1) or the derivative of the zika virus NS1, a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B, a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3, a polynucleotide encoding a zika virus non- structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, or any combination of two or more thereof. The polynucleotide of interest plasmid or third polynucleotide may comprise a polynucleotide of interest.

[0165] In some embodiment, the producer cell is a Vero E6, HEK, HEK 293T, HEK 293TT, FreeStyle™ 293 -F Cell, HEK-293.2sus or C6 / 36 cell.

[0166] In some embodiments, the zika virus-like particle is not replication competent. In some embodiments, the zika virus-like particle is replication incompetent without lowering the vector titer or impairing expression of the polynucleotide of interest. In some embodiments, the zika virus-like particle has decreased insertional mutagenesis. In some embodiments, the zika viruslike particle has decreased immune response. In some embodiments, the zika virus-like particle has tropism for a cell of the central nervous system and / or the ocular system.

[0167] In some embodiments, the zika virus-like particle comprise: a polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the polynucleotide does not encode one or more viral non-structural proteins and one or more viral structural proteins. In some embodiments, the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, and zika virus NS5 or a derivative of the zika virus NS5; and wherein the one or more viral structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, and zika virus viral envelope protein (E) or a derivative of the zika virus E. In some embodiments, the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, or zika virus NS5 or a derivative of the zika virus NS5, or a combination of two or morethereof. In some embodiments, the polynucleotide encodes a zika virus NS4a comprising a sequence at least 90% identical to SEQ ID NO: 2929 or 2946. In some embodiments, the polynucleotide encodes a zika virus NS4a comprising a pTMS3. In some embodiments, the polynucleotide does not encode a zika virus NS4a comprising a pTMSl and a pTMS2. In some embodiments, the polynucleotide does not encode a full length zika virus NS4a. In some embodiments, the full length zika virus NS4a comprises a sequence at least 90% identical to SEQ ID NO: 10 or 2942. In some embodiments, the polynucleotide encodes a NS4b comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948. In some embodiments, the polynucleotide encodes a NS4b comprising a N-terminal disordered region. In some embodiments, the polynucleotide does not encode a zika virus NS4b comprising a pTMSl, a pTMS2, a pTMS3, a pTMS4, and a pTMS5. In some embodiments, the polynucleotide does not encode a full length zika virus NS4b. In some embodiments, the full length zika virus NS4b comprises a sequence at least 90% identical to SEQ ID NO: 12 or 2944. In some embodiments, the polynucleotide encodes a zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947. In some embodiments, the one or more structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, zika virus viral envelope protein (E) or a derivative of the zika virus E, or a combination thereof. In some embodiments, the polynucleotide encodes a zika virus capsid protein (C) comprising a sequence at least 90% identical to any one of SEQ ID NOS: 2908-2917. In some embodiments, the polynucleotide does not encode a full length zika virus C. In some embodiments, the full length zika virus C comprises a sequence at least 90% identical to SEQ ID NO: 3 or 2901. In some embodiments, the zika virus-like particle comprises the one or more viral non-structural proteins and the one or more viral structural proteins, optionally wherein the one or more structural proteins are assembled into a zika virus-like shell, and further optionally wherein the zika viruslike particle is not replication competent.Methods

[0168] In another aspect, provided herein is a method of delivering an polynucleotide of interest to a target cell, the method comprising applying to the target cell a zika virus-like particle described herein comprising the polynucleotide of interest.

[0169] In some embodiments, the target cell is a neuron or a non-neuron cell. In specific embodiments, the neuron is an oligodendrocyte, microglia, or astrocyte. In another specific embodiments, the non-neuron cell is a prostate epithelial cell, a urethra epithelial cell, a Sertoli cell, a Leydig cell, a spermatogonium cell or a retinal cell.

[0170] In some embodiments, the method is carried out in vitro, ex vivo, or in vivo.

[0171] In some embodiments, the target cell transiently expresses the polynucleotide of interest after delivery. In other embodiments, the target cell persistently expresses the polynucleotide of interest after delivery.

[0172] In another aspect, provided herein is a method of treating Alzheimer’s disease in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding a brain-derived neurotrophin factor (BDNF).

[0173] In another aspect, provided herein is a method of treating Alzheimer’s disease in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide targeting beta-site amyloid precursor protein cleaving enzyme 1 (BACE1).

[0174] In another aspect, provided herein is a method of treating Alzheimer’s disease in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide targeting amyloid precursor protein (APP).

[0175] In another aspect, provided herein is a method of treating Alzheimer’s disease in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide targeting Tau.

[0176] In another aspect, provided herein is a method of treating autism in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding SH3 and multiple ankyrin repeat domains protein (SHANK).

[0177] In yet another aspect, provided herein is a method of treating Parkinson’s disease in a subject in need thereof, the method comprising administering the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding aromatic L- amino acid decarboxylase (AADC).

[0178] In another aspect, provided herein is a method of treating Parkinson’s disease in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding a human aromatic L-amino acid decarboxylase.

[0179] In another aspect, provided herein is a method of treating Parkinson’s disease in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding glial cell line- derived neurotrophic factor.

[0180] In another aspect, provided herein is a method of treating / managing Down’s Syndrome in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide targets the additional copy of HSA21.

[0181] In another aspect, provided herein is a method of treating choroideremia-blindness in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding rab-escort protein 1 (REP1).

[0182] In another aspect, provided herein is a method of treating leber congenital amaurosis in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding an RPE65.

[0183] In another aspect, provided herein is a method of treating Parkinson’s disease in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding a glutamic acid decarboxylase.

[0184] In another aspect, provided herein is a method of treating ornithine transcarbamylase (OTC) deficiency in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding ornithine transcarbamylase.

[0185] In another aspect, provided herein is a method of treating multiple sclerosis in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding an interferon- beta-la and lb.

[0186] In another aspect, provided herein is a method of treating Pompe Disease in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding an acid alphaglucosidase.

[0187] In another aspect, provided herein is a method of treating depression in a subject in need thereof, and the method comprises administering to the subject an effective amount of the zika virus-like particle as described herein comprising a polynucleotide encoding a selective serotonin reuptake inhibitor.

[0188] In another aspect, provided herein is a method of triggering or boosting an immune response in a subject, the method comprising administering to the subject an effective amount ofa peptide produced from the nucleic acid composition as described herein comprising an antigen or an antigenic epitope thereof.

[0189] In some embodiments, the peptide is produced from a transcript produced from an polynucleotide of interest from a zika virus-like particle.

[0190] In some embodiments, the administrating is performed intramuscularly.

[0191] In some embodiments, the antigen or an antigenic epitope thereof is from a pathogen.

[0192] In some embodiments, the pathogen is a virus. In specific embodiments, the virus is a human SARS coronavirus, an influenza A virus, an influenza B virus, an influenza C virus an ebolavirus, a hepatitis B virus, a hepatitis C virus, a herpes simplex virus, a human immunodeficiency virus (HIV), a human papillomavirus (HPV-6, HPV-11), a measles virus, a rabies virus, a poliovirus, or a yellow fever virus.

[0193] In some embodiments, the pathogen is a bacteria. In specific embodiments, the bacteria is Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella, henselae, Bartonella quintana, Bifidobacterium Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, Cardiobacterium hominis, Campylobacter fetus, Chlamydia pneumonia, Chlymydia trahomatis, Clostridium difficile, Cyanobacteria, Eikennella corrodens, Enterobacter, Enterococcus faccium, Escherichia coli, Escherichia coli 0157, Franceilla tularensis, Fusobacterium nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzae, Helicobacter pylori, Kingella kingae, Klebsiella pneumonia, Legionella bacteria, Legionella pneumophila serogroup 1, Leptospria, Morganella morganii, Neisseria gonorrhoeae, Neisseria meningitidis, Proteus mirabilis, Proteus vulgaris, Proteus myxofaciens, Providencia rettgeri, Providencia alcalifaciens, Providencia stuartii, Pseudomonas aeruginosa, Pseudomonas paucimobilis, Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas acidovorans, Rickettsiae, Salmonella enterica, Salmonella typhi, Salmonella paratyphi types A, B typhus, Salmonella, dublin, Salmonella arizonae, Salmonella choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonnei, Treponema, Stenotrophomonas maltophilia, Vibrio cholerae, Vibrio mimicus, Vibrio alginolyticus, Vibrio hollisae, Vibrio parahaemolyticus, Vibrio vulnificus, Yersinia pestitis, Actinomycetes, Bacillus anthracis, Bacillus subtilis, Clostridium tetani, Clostridium, perfmgens, Clostridium botulinum, Clostridium tetani. Corynebacterium diphtheriae, Enterococcus faecalis, Enterococcus faecium, Erysipelothrix ruhsiopathiae, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium tuberculosis, Mycoplasma, Nocardia, Propionibacerium, Pseudomonas aeruginosa, Pneumococci, Staphylococcus aureus, Staphylococcus epidermidis, methicillin resistant Staphylococcus aureus(MRSA), vancomycin resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcus saprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutants.

[0194] In some embodiments, the pathogen is a fungus, an amoeba, or a parasite. In specific embodiments, the fungus, the amoeba, or the parasite is Acanthamoeba spp, American tryppanosomiasis, Balamuthia mandnillanis, Babesia divergenes, Babesia bigemina, Babesia equi, Babesia microfti, Babesia duncani, Balantidium coli, Blastocystis spp Cryptosporidium spp, Cyclospora cayetanensis, dientamoeba fragilis, Diphyllobothrium latum, Leishmania amazonesis, Naegleria fowderi, Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale curtisi, Plasmodium malariae, Rhinosporidium seeberi, Sarcocystis bovihominis, Sarcocystiss suihominis, Toxoplasma gondii, Trichmonas vaginalis, Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0195] In some embodiments, the administrating is via systemic delivery.

[0196] In some embodiments, the administrating is performed intravenously and / or intratumorly.

[0197] In some embodiments, the administrating targets the cerebral spinal fluid in intracerebroventricular, cistema magna, subpial, and / or intrathecal. In other embodiments, the administrating is not performed intraparenchymally.Dosing and treatment regimens

[0198] Administration frequencies for a pharmaceutical composition comprising the zika viruslike particle provided herein may vary based on the method being practiced, the physical characteristics of the subject, the severity of the disease, disease type, and the formulation and the means used to administer the composition.

[0199] The amount of the zika virus-like particle provided herein varies depending upon factors such as the particular virus, disease condition and its severity, the identity (e.g., weight, sex) of the subject in need of treatment.Pharmaceutical compositions and formulations of the zika virus-like particle

[0200] Provided herein are zika virus-like particles and nucleic acid compositions formulated into pharmaceutical compositions.

[0201] Pharmaceutical compositions are formulated in a conventional manner using one or more pharmaceutically acceptable inactive ingredients that facilitate processing of the active agent into preparations that can be used pharmaceutically.

[0202] In certain embodiments, a pharmaceutical composition of this disclosure can comprise an effective amount of a nucleic acid composition or virus-like particle, disclosed herein, combined with a pharmaceutically acceptable carrier. "Pharmaceutically acceptable," as used herein,includes any carrier which does not interfere with the effectiveness of the biological activity of the active ingredients and / or that is not toxic to the patient to whom it is administered.Treatment in cancer

[0203] An aspect of this disclosure relates to methods of cancer treatment comprising administering a zika virus-like particle as described herein. In example embodiments, the zika virus-like particle comprises a polynucleotide of interest. The polynucleotide of interest may comprise or encode a therapeutic agent (e.g., nucleic acid or peptide) that is effective for treating cancer. In some embodiments, the polynucleotide of interest is delivered to cancerous cells. For instance, the zika virus-like particle has tropism for a cancer cell. In some embodiments, the zika virus-like particle has tropism for a cell undergoing cell division. In some embodiments, the polynucleotide of interest is not delivered to non-cancerous cells.

[0204] In some embodiments, the zika virus-like particle has tropism for a cell in the central nervous system. In some embodiments, the zika virus-like particle has tropism for a cell in the ocular system.

[0205] In some embodiments, the zika virus-like particle impairs cell division, impairs cell growth, and / or induces cell death of cancerous cells. In some embodiments, the zika virus-like particle does not impair cell division, cell growth, and / or induce cell death in non-cancerous cells.

[0206] Cancer is a class of diseases that may be characterized by uncontrolled cellular growth. Cancer includes all types of hyperproliferative growth, hyperplastic growth, neoplastic growth, cancerous growths or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues, or organs, irrespective of histopathologic type or stage of invasiveness. Types of cancer include, but are not limited to, solid tumors and hematological tumors at any stage of the disease, with or without metastases.

[0207] In some embodiments, the cancer is a CNS cancer. In some embodiments, the cancer is brain cancer. Non-limiting examples of cancers treatable with a zika virus-like particle provided herein include adenoma, angio-sarcoma, astrocytoma, epithelial carcinoma, germinoma, glioblastoma, glioma, hamartoma, hemangioendothelioma, hemangiosarcoma, hematoma, hepato-blastoma, leukemia, lymphoma, medulloblastoma, melanoma, neuroblastoma, osteosarcoma, retinoblastoma, rhabdomyosarcoma, sarcoma, and teratoma. Glioma refers to a tumor originating in the neuroglia of the brain or spinal cord. Gliomas are derived from the glial cell types such as astrocytes and oligodendrocytes, thus gliomas include astrocytomas and oligodendrogliomas, as well as anaplastic gliomas, glioblastomas, and ependymomas. Additional brain tumors include meningiomas, ependymomas, pineal region tumors, choroid plexus tumors,neuroepithelial tumors, embryonal tumors, peripheral neuroblastic tumors, tumors of cranial nerves, tumors of the hemopoietic system, germ cell tumors, and tumors of the sellar region.

[0208] In some specific embodiments, the brain cancer is an astrocytoma, an oligodendroglioma, an ependymoma, a meningioma, a schwannoma, a craniopharyngioma, a germinoma, a pineocytoma, or a combination thereof.

[0209] In some cases the zika virus-like particle interacts with an Integrin Subunit Alpha V (ITGAV) receptor. Non-limiting example tumor cells that express this receptor include, cholangio (bile duct), esophagus, kidney, liver, lymphonodus, breast, stomach, pancreatic, head and neck, Thymoma, thyroid, and colon tumor cells. As such, in some specific embodiments, zika virus-like particles described herein are used to treat any one of the following cancers: cholangio (bile duct), esophagus, kidney, liver, lymphonodus or cancer of the lymph node, breast, stomach, pancreatic, head and neck, Thymoma, thyroid, and colon cancer.

[0210] Methods of treatment may include contacting a cancerous cell with an effective amount of a zika virus-like particle described herein. The contacting may be carried out in vitro (e.g., in biochemical and / or cellular assays), in vivo in a non-human animal, and in vivo in mammals, including humans. In some cases, contacting involves bringing a cancer cell and a composition comprising the zika virus-like particle described herein into sufficient proximity such that the composition exerts an effect on the cancer cell. Contacting includes physical interaction between the composition and a cancer cell, as well as interactions that do not require physical interaction. Contacting includes administering an effective amount of a composition to a subject having the cancer cell such that the composition impairs cancer cell growth, division and / or induces cancer cell death. In some embodiments, contacting by administration includes intravenous administration, intraperitoneal administration, intra-tumoral, intramuscular administration, intracoronary administration, intraarterial administration, subcutaneous administration, transdermal delivery, intratracheal administration, subcutaneous administration, intraarticular administration, intraventricular administration, inhalation, intracerebral, nasal, oral, pulmonary administration, impregnation of a catheter, and direct injection into a tissue or tumor of the subject. In some embodiments, contacting by administration includes intraperitoneal administration. In some embodiments, contacting by administration includes intra-tumoral injection.

[0211] Subjects include animals, such as humans, other higher primates, lower primates, and animals of veterinary importance, such as dogs, cats, horses, sheep, goats, and cattle and the like. Subjects also include animals for use in studies, for example, mice, rats and other rodents.

[0212] Method for treating cancer may include administration of an effective amount of a zika virus-like particle described herein to a subject. An effective amount or therapeutically effective amount is a quantity sufficient to achieve a desired therapeutic and / or prophylactic effect, e.g., an amount which results in the prevention of, or a decrease in, the symptoms associated with a cancer. The amount administered to the subject may depend on the type and severity of the cancer and on the characteristics of the individual, such as general health, age, sex, body weight and tolerance to drugs. In some embodiments, an effective amount is sufficient to slow, inhibit and / or reduce the growth or size of a tumor, and includes the eradication of the tumor in certain instances. In some embodiments, an effective amount is administered to a subject having cancer in an amount sufficient to attenuate or halt division of a cancer cell.

[0213] Treatment of cancer includes amelioration, cure, and / or maintenance of a cure (i.e. the prevention or delay of relapse) and / or its associated symptoms. For example, a subject is successfully treated for a cancer if after receiving a therapeutic amount of a zika virus-like particle described herein, the subject shows observable and / or measurable reduction in or absence of one or more signs and symptoms of the cancer, e.g., reduction in the number of cancer cells or absence of the cancer cells; reduction in the tumor size; inhibition (i.e., slow to some extent and preferably stop) of tumor metastasis; inhibition, to some extent, of tumor growth; increase in length of remission, and / or relief to some extent, of one or more of the symptoms associated with the specific cancer; reduced morbidity and mortality, and improvement in quality of life issues. Treatment also includes preventing the cancer from becoming worse, slowing the rate of progression, and / or preventing the cancer from re-occurrence after initial elimination.

[0214] In some embodiments, a therapeutic benefit is achieved after administration of a zika virus-like particle described herein. A therapeutic benefit includes anything that promotes or enhances the well-being of the subject with respect to the medical treatment of his / her condition, which includes treatment of pre-cancer, cancer, and hyperproliferative diseases. For example, extension of the subject's life by any period of time, decrease or delay in the neoplastic development of the disease, decrease in hyperproliferation, reduction in tumor growth, delay of metastases, reduction in cancer cell or tumor cell proliferation rate, and a decrease in pain to the subject that can be attributed to the subject's condition.

[0215] In some instances, tumor-selective targeting of a zika virus-like particle as described herein can be measured through use of a reporter gene, such as a luciferase gene. In some embodiments, the luciferase gene can be included in the zika virus-like particle, and a tumor cell can be contacted with the particle. Bioluminescence in tumor cells can be measured to monitor tumor-selective targeting.Kits of the zika virus-like particle

[0216] In one aspect of the disclosure, provided herein are kits which include one or more reagents or devices for the performance of the methods disclosed herein. In some embodiments, the kit comprises the zika virus-like particle or nucleic acid composition provided herein. In some embodiments, the kit comprises a means to administrate the zika virus-like particle or nucleic acid composition provided herein.

[0217] In some embodiments, the kit comprises suitable instructions in order to perform the methods of the kit. The instructions may provide information of performing any of the methods disclosed herein, whether or not the methods may be performed using only the reagents provided in the kit. The kit and instructions may require additional reagents or systems.

[0218] For use in the therapeutic applications described herein, kits and articles of manufacture are also described herein. In some embodiments, such kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) including one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers can be formed from a variety of materials such as glass or plastic. The articles of manufacture provided herein contain packaging materials. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material suitable for a selected formulation and intended mode of administration and treatment. The container(s) optionally have a sterile access port (for example the container is an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). Such kits optionally comprise a composition with an identifying description or label or instructions relating to its use in the methods described herein.

[0219] A kit will typically include one or more additional containers, each with one or more of various materials (such as reagents, optionally in concentrated form, and / or devices) desirable from a commercial and user standpoint for use of the zika virus-like particle or nucleic acid composition described herein. Non-limiting examples of such materials include, but not limited to, buffers, diluents, filters, needles, syringes, carrier, package, container, vial and / or tube labels listing contents and / or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included.

[0220] In some embodiments, a label is on or associated with the container. A label can be on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself; a label can be associated with a container when it is presentwithin a receptacle or carrier that also holds the container, e.g., as a package insert. A label can be used to indicate that the contents are to be used for a specific therapeutic application. The label can also indicate directions for use of the contents, such as in the methods described herein.

[0221] In certain embodiments, a pharmaceutical composition comprising the zika virus-like particle provided herein and optional additional active agent is presented in a pack or dispenser device which can contain one or more unit dosage forms. The pack can for example contain metal or plastic foil, such as a blister pack. The pack or dispenser device can be accompanied by instructions for administration. The pack or dispenser can also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, can be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. Compositions containing the zika virus-like particle described herein formulated in a compatible pharmaceutical carrier can also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.Table 1A Non-limiting example Zika sequencesTable IB Non-limiting example Zika nucleotide sequencesTable 1C Non-limiting example exogenous 5’ UTR sequencesTable ID Non-limiting example exogenous 3’ UTR sequencesTable IE Non-limiting examples polynucleotides of interestEMBODIMENTS

[0222] Embodiment 1. A zika virus-like particle, comprising: (a) a zika virus structural protein, (b) optionally a zika virus non- structural (NS) protein, and (c) a polynucleotide of interest, wherein the zika virus-like particle is not replication competent; optionally wherein the zika virus-like particle is generated from a composition comprising a first polynucleotide encoding the zika virus structural protein, a second polynucleotide encoding the zika virus non-structural (NS) protein, and a third polynucleotide comprising the polynucleotide of interest.

[0223] Embodiment 2. The zika virus-like particle or the composition of embodiment 1, wherein the zika virus structural protein comprises: (a) a zika virus capsid protein (C), or a derivative of a zika virus C (b) a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M (c) a zika virus viral envelope protein (E) or a derivative of a zika virus E, or (d) a combination of two or more selected from (a) to (c).

[0224] Embodiment 3. The zika virus-like particle or the composition of embodiments 1-2, wherein the zika virus non-structural protein comprises: (a) zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1, (b) a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (c) a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (d) a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B, (e) a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3, (f) a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, (g) a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, (h) a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, (i) a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, (j) a poly A tail, (k) or a zika virus viral envelope protein (E) or a derivative of a zika virus E (1) a combination of two or more selected from (a) to (k).

[0225] Embodiment 4. The zika virus-like particle or the composition of any one of embodiments 1-3, wherein the third polynucleotide further comprises: (a) a polynucleotide encoding a zika virus 5’ UTR or a derivative of the zika virus 5’ UTR, (b) a polynucleotide encoding a zika virus 3’ UTR or a derivative of the zika virus 3’ UTR, (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C, (d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M, (e) a polynucleotide encoding a zika virus viral envelope protein E or a derivative of a zika virus E, (f) a polynucleotide of interest, or (g) a combination of two or more selected from (a) to (f).

[0226] Embodiment 5. The zika virus-like particle or the composition of any one of embodiments 1-4, wherein the zika virus-like particle is replication incompetent without impairing expression of the polynucleotide of interest.

[0227] Embodiment 6. The zika virus-like particle or the composition of any one of embodiments 1-5, wherein the zika virus-like particle has decreased immune response as compared to a wild-type zika virus.

[0228] Embodiment 7. The zika virus-like particle or the composition of any one of embodiments 1-6, wherein the zika virus-like particle has tropism for central nervous system or ocular system cells.

[0229] Embodiment 8. The zika virus-like particle or the composition of any one of embodiments 1-7, further comprising a pharmaceutically acceptable salt or derivative thereof.

[0230] Embodiment 9. The zika virus-like particle or the composition of any one of embodiments 1-8, wherein the zika virus structural protein is expressed from the first polynucleotide and the first polynucleotide is a structural protein plasmid.

[0231] Embodiment 10. The zika virus-like particle or the composition of any one of embodiments 1-9, wherein the first polynucleotide or the structural protein plasmid comprises: (a) a polynucleotide encoding a 5’ UTR, (b) a polynucleotide encoding a 3’ UTR, (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C; (d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; (e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E, or (f) a combination of two or more selected from (a) to (e).

[0232] Embodiment 11. A structural protein plasmid comprising: (a) a polynucleotide encoding a 5’ UTR, (b) a polynucleotide encoding a 3’ UTR, (c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C; (d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; (e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E, or (f) a combination of two or more selected from (a) to (e).

[0233] Embodiment 12. The zika virus-like particle or the composition of embodiment 10, or the structural protein plasmid of embodiment 11, wherein the polynucleotide encoding the 5’ UTR comprises a 5’ UTR from a zika virus of Table IB, or a 5’ UTR of Table 1C, Table IF, Table 3 or Table 4.

[0234] Embodiment 13. The zika virus-like particle or the composition of embodiment 10, or the structural protein plasmid of embodiment 11, wherein the polynucleotide encoding the 5’ UTR comprises a polynucleotide at least 80% identical to a 5’ UTR from a zika virus of Table IB or a 5’ UTR of Table 1C, Table IF, Table 3 or Table 4.

[0235] Embodiment 14. The zika virus-like particle or the composition of any one of embodiments 10 or 12-13, or the structural protein plasmid of any one of embodiments 11-13, wherein the polynucleotide encoding the 3’ UTR comprises a 3’ UTR from a zika virus of Table IB, or a 3’ UTR of Table ID, Table IF, Table 3 or Table 4.

[0236] Embodiment 15. The zika virus-like particle or the composition of any one of embodiments 10 or 12-13, or the structural protein plasmid of any one of embodiments 11-13, wherein the polynucleotide encoding the 3’ UTR comprises a polynucleotide at least 80% identical to a3’ UTR from a zika virus of Table IB, or a 3’ UTR of Table ID, Table IF, Table 3 or Table 4.

[0237] Embodiment 16. The zika virus-like particle or the composition of any one of embodiments 10 or 12-15, or the structural protein plasmid of any one of embodiments 11-15, wherein the polynucleotide encoding the derivative of the zika virus C comprises at least one-substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus C.

[0238] Embodiment 17. The zika virus-like particle or the composition of any one of embodiments 10 or 12-16, or the structural protein plasmid of any one of embodiments 11-16, wherein the zika virus C or the derivative of the zika virus C comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus C from Table 1 A, Table IF, Table 2, or Table 3, or the zika virus C from zika virus of Table IB.

[0239] Embodiment 18. The zika virus-like particle or the composition of any one of embodiments 10 or 12-17, or the structural protein plasmid of embodiments 11-17, wherein the polynucleotide encoding the zika virus C or the derivative of the zika virus C comprises a sequence at least 80% identical to a zika virus C sequence of Table 1A, Table IF, Table 2, or Table 3, or the zika virus C from a zika virus of Table IB.

[0240] Embodiment 19. The zika virus-like particle or the composition of any one of embodiments 10 or 12-18, or the structural protein plasmid of any one of embodiments 11-18, wherein the polynucleotide encoding the derivative of the zika virus prM / M comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus prM / M.

[0241] Embodiment 20. The zika virus-like particle or the composition of any one of embodiments 10-19, or the structural protein plasmid of any one of embodiments 11-19, wherein the zika virus prM / M protein or the derivative of the zika virus prM / M comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus prM / M from of Table 1A, Table IB or Table 3.

[0242] Embodiment 21. The zika virus-like particle or the composition of any one of embodiments 10-20, or the structural protein plasmid of any one of embodiments 11-20, wherein the polynucleotide encoding the zika virus prM / M or the derivative of the zika virus prM / M comprises a sequence at least 80% identical to a prM / M sequence of Table 1A, Table IF or Table 3, or the zika virus prM / M sequence from a zika virus of Table IB.

[0243] Embodiment 22. The zika virus-like particle or the composition of any one of embodiments 10-21, or the structural protein plasmid of any one of embodiments 11-21, wherein the polynucleotide encoding the derivative of the zika virus E comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus E.

[0244] Embodiment 23. The zika virus-like particle or the composition of any one of embodiments 10-22, or the structural protein plasmid of any one of embodiments 11-22, whereinthe zika virus E protein or the derivative of the zika virus E comprises an amino acid sequence at least 80% identical to a sequence of Table 1A, Table IB or Table 3.

[0245] Embodiment 24. The zika virus-like particle or the composition of any one of embodiments 10-23, or the structural protein plasmid of embodiments 11-23, wherein the polynucleotide encoding the zika virus E or the derivative of the zika virus E comprises a sequence at least 80% identical to an amino acid sequence encoded by a zika virus E from Table 1A, Table IB, Table F or Table 3.

[0246] Embodiment 25. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-24, or the structural protein plasmid of any one of embodiments 11-24, wherein the first polynucleotide or the structural protein plasmid does not further comprise: (a) zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1, (b) a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (c) a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (d) a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B, (e) a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3, (f) a zika virus non- structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, (g) a zika virus non- structural protein 2K (2K) or the derivative of the zika virus 2K, (h) a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, (i) a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, (k) a poly A tail, (1) or a zika virus viral envelope protein (E) or a derivative of a zika virus E or (m) a combination of two or more selected from (a) to (1).

[0247] Embodiment 26. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-25, or the structural protein plasmid of any one of embodiments 11-25, further comprising: (a) zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1, (b) a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (c) a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (d) a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B, (e) a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3, (f) a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, (g) a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, (h) a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, (i) a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, (k) a poly A tail, (1) or a zika virus viral envelope protein (E) or a derivative of a zika virus E or (m) a combination of two or more selected from (a) to (1).

[0248] Embodiment 27. The zika virus-like particle or the composition of embodiment 25 or embodiment 26, or the structural protein plasmid of embodiment 25 or embodiment 26, wherein the NS 1 is a zika virus NS 1, the NS2A is a zika virus NS2A, the NS2B is a zika virus NS2B, the NS3 is a zika virus NS3, the NS4A is a zika virus NS4A, the NS4B is a zika virus NS4B, or the NS5 is a zika virus NS5.

[0249] Embodiment 28. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-27, or the structural protein plasmid of any one of embodiments 11-27, wherein the structural protein is a structural protein from an African zika virus, Asian zika virus, or Brazilian zika virus, or a combination of two or more.

[0250] Embodiment 29. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-28, or the structural protein plasmid of any one of embodiments 11-28, wherein the non-structural protein is a non-structural protein from an African zika virus, Asian zika virus, or Brazilian zika virus, or a combination of two or more.

[0251] Embodiment 30. The zika virus-like particle or the composition of any one of embodiments 28-29, or the structural protein plasmid of any one of embodiments 28-29, wherein the African zika virus is African MR766 strain.

[0252] Embodiment 31. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-30, or the structural protein plasmid of any one of embodiments 11-30, wherein the first polynucleotide or the structural protein plasmid further comprises one or more expression control elements in operable linkage that confers expression of the first polynucleotide or the structural protein plasmid in vitro or in vivo.

[0253] Embodiment 32. The zika virus-like particle or the composition of embodiment 31, or the structural protein plasmid of embodiment 31, wherein the one or more expression control elements is a promoter that drives expression of the first polynucleotide or the structural protein plasmid in vitro.

[0254] Embodiment 33. The zika virus-like particle or the composition of embodiment 32, or the structural protein plasmid of embodiment 32, wherein the promoter is a phage promoter.

[0255] Embodiment 34. The zika virus-like particle or the composition of embodiment 33, or the structural protein plasmid of embodiment 33, wherein the phage promoter is T7, T3, or SP6.

[0256] Embodiment 35. The zika virus-like particle or the composition of embodiment 32, or the structural protein plasmid of embodiment 32, wherein the promoter is a CMV promoter, a SV40 promoter, a eukaryotic promoter, a elongation factor EF-la promoter, a chicken beta-actin promoter, or a phosphoglycerate kinase promoter.

[0257] Embodiment 36. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-35, or the structural protein plasmid of embodiments 11- 35, wherein the first polynucleotide or the structural protein plasmid further comprises a eukaryotic enhancer.

[0258] Embodiment 37. The zika virus-like particle or the composition of embodiment 36, or the structural protein plasmid of embodiment 36, wherein the eukaryotic enhancer is a CMV enhancer or a SV40 enhancer.

[0259] Embodiment 38. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-37, or the structural protein plasmid of any one of embodiments 11-37, wherein the first polynucleotide or the structural protein plasmid further comprises an RNA polymerase promoter.

[0260] Embodiment 39. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-38, wherein the zika virus non- structural protein is expressed from the second polynucleotide and the second polynucleotide is a non- structural protein plasmid.

[0261] Embodiment 40. The zika virus-like particle or the composition of any one of embodiments 1-10 or embodiments 12-39, wherein the second polynucleotide or the non- structural protein plasmid comprises: (a) a polynucleotide encoding a 5’ UTR, (b) a polynucleotide encoding a 3’ UTR, (c) a polynucleotide encoding a zika virus non-structural protein 1 (NS1) or the derivative of the zika virus NS1, (d) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B, (g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3, (h) a polynucleotide encoding a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, (i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, (j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, (k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, (k) a poly A tail, (1) or a zika virus viral envelope protein (E) or a derivative of a zika virus E or (m) a combination of two or more selected from (a) to (1).

[0262] Embodiment 41. A non-structural protein plasmid comprising: (a) a polynucleotide encoding a 5’ UTR, (b) a polynucleotide encoding a 3’ UTR, (c) a polynucleotide encoding azika virus non- structural protein 1 (NS1) or the derivative of the zika virus NS1, (d) a polynucleotide encoding a zika virus non- structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (e) a polynucleotide encoding a zika virus non- structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, (f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B, (g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3, (h) a polynucleotide encoding a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, (i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, (j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, (k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS5, (k) a poly A tail, (1) or a zika virus viral envelope protein (E) or a derivative of a zika virus E or (m) a combination of two or more selected from (a) to (1).

[0263] Embodiment 42. The zika virus-like particle or the composition of embodiment 40, or the non-structural protein plasmid of embodiment 41, wherein the polynucleotide encoding the 5’ UTR comprises a 5’ UTR from a zika virus of Table IB, or a 5’ UTR of Table 1C, Table IF, Table 3, or Table 4.

[0264] Embodiment 43. The zika virus-like particle or the composition of embodiment 40, or the non-structural protein plasmid of embodiment 41, wherein the polynucleotide encoding the 5’ UTR comprises a polynucleotide at least 80% identical to a 5’ UTR from a zika virus of Table IB, or a 5’ UTR of Table 1C, Table IF, Table 3 or Table 4.

[0265] Embodiment 44. The zika virus-like particle or the composition of any one of embodiments 40 or 42-43, or the non-structural protein plasmid of any one of embodiments 41-43, wherein the polynucleotide encoding the 3’ UTR comprises a 3’ UTR from a zika virus of Table IB, or a 3’ UTR of Table ID, Table IF, Table 3 or Table 4.

[0266] Embodiment 45. The zika virus-like particle or the composition of any one of embodiments 40 or 42-44, or the non-structural protein plasmid of any one of embodiments 41-44, wherein the polynucleotide encoding the 3’ UTR comprises a polynucleotide at least 80% identical to a 3’ UTR from a zika virus of Table IB, or a 3’ UTR of Table ID, Table IF, Table 3 or Table 4.

[0267] Embodiment 46. The zika virus-like particle or the composition of any one of embodiments 40 or 42-45, or the non-structural protein plasmid of any one of embodiments 41-45, wherein the polynucleotide encoding the derivative of the zika virus NS1 comprises at leastone substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus NS1.

[0268] Embodiment 47. The zika virus-like particle or the composition of any one of embodiments 40 or 42-46, or the non-structural protein plasmid of any one of embodiments 41-46, wherein the zika virus NS1 or the derivative of the zika virus NS1 comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus NS1 from Table IB, or a NS 1 of Table 4.

[0269] Embodiment 48. The zika virus-like particle or the composition of any one of embodiments 40 or 42-47, or the non-structural protein plasmid of any one of embodiments 41-47, wherein the polynucleotide encoding the zika virus NS1 or the derivative of the zika virus NS 1 comprises a sequence at least 80% identical to a NS 1 sequence of Table 4, or the zika virus NS1 from a zika virus of Table IB.

[0270] Embodiment 49. The zika virus-like particle or the composition of any one of embodiments 40 or 42-48, or the non-structural protein plasmid of any one embodiments 41-48, wherein the polynucleotide encoding the derivative of the zika virus NS2A comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus NS2A.

[0271] Embodiment 50. The zika virus-like particle or the composition of any one of embodiments 40 or 42-49, or the non-structural protein plasmid of any one of embodiments 41-49, wherein the zika virus NS2A protein or the derivative of the zika virus NS2A comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus NS1 from Table 4, or a NS2A from a zika virus of Table IB.

[0272] Embodiment 51. The zika virus-like particle or the composition of any one of embodiments 40 or 42-50, or the non-structural protein plasmid of any one of embodiments 41-50, wherein the polynucleotide encoding the zika virus NS2A or the derivative of the zika virus NS2A comprises a sequence at least 80% identical to a NS2A sequence of Table 4, or a NS2A from a zika virus of Table IB.

[0273] Embodiment 52. The zika virus-like particle or the composition of any one of embodiments 40 or 42-51, or the non-structural protein plasmid of any one of embodiments 41-51, wherein the polynucleotide encoding the derivative of the zika virus NS2B comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus NS2B.

[0274] Embodiment 53. The zika virus-like particle or the composition of any one of embodiments 40 or 42-52, or the non-structural protein plasmid of any one of embodiments 41-52, wherein the zika virus NS2B protein or the derivative of the zika virus NS2B comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus NS1 from Table 4, or a NS2B from a zika virus of Table IB.

[0275] Embodiment 54. The zika virus-like particle or the composition of any one of embodiments 40 or 42-53, or the non-structural protein plasmid of any one of embodiments 41-53, wherein the polynucleotide encoding the zika virus NS2B or the derivative of the zika virus NS2B comprises a sequence at least 80% identical to a NS2B of Table 4, or a NS2B from a zika virus of Table IB.

[0276] Embodiment 55. The zika virus-like particle or the composition of any one of embodiments 40 or 42-54, or the non-structural protein plasmid of any one of embodiments 41-54, wherein the polynucleotide encoding the derivative of the zika virus NS3 comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus NS3.

[0277] Embodiment 56. The zika virus-like particle or the composition of any one of embodiments 40 or 42-55, or the non-structural protein plasmid of any one of embodiments 41-55, wherein the zika virus NS3 or the derivative of the zika virus NS3 comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus NS3 from Table 4, or a NS3 from a zika virus of Table IB.

[0278] Embodiment 57. The zika virus-like particle or the composition of any one of embodiments 40 or 42-56, or the non-structural protein plasmid of any one of embodiments 41-56, wherein the polynucleotide encoding the zika virus NS3 or the derivative of the zika virus NS3 comprises a sequence at least 80% identical to a NS3 from Table 4, or a NS3 from a zika virus of Table IB.

[0279] Embodiment 58. The zika virus-like particle or the composition of any one of embodiments 40 or 42-57, or the non-structural protein plasmid of any one of embodiments 41-57, wherein the polynucleotide encoding the derivative of the zika virus NS4A comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus NS4A.

[0280] Embodiment 59. The zika virus-like particle or the composition of any one of embodiments 40 or 42-58, or the non-structural protein plasmid of any one of embodiments 41-58, wherein the zika virus NS4A protein or the derivative of the zika virus NS4A comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus NS4A from Table 4, or a NS4A from a zika virus of Table IB.

[0281] Embodiment 60. The zika virus-like particle or the composition of any one of embodiments 40 or 42-59, or the non-structural protein plasmid of any one of embodiments 41-59, wherein the polynucleotide encoding the zika virus NS4A or the derivative of the zika virus NS4A comprises a sequence at least 80% identical to a NS4A from Table 4, or a NS4A from a zika virus of Table IB.

[0282] Embodiment 61. The zika virus-like particle or the composition of any one of embodiments 40 or 42-60, or the non-structural protein plasmid of any one of embodiments 41-60, wherein the polynucleotide encoding the derivative of the zika virus 2K comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus 2K.

[0283] Embodiment 62. The zika virus-like particle or the composition of any one of embodiments 40 or 42-61, or the non-structural protein plasmid of any one of embodiments 41-61, wherein the zika virus 2K protein or the derivative of the zika virus 2K comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus 2K from Table 4, or a 2K from a zika virus of Table IB.

[0284] Embodiment 63. The zika virus-like particle or the composition of any one of embodiments 40 or 42-62, or the non-structural protein plasmid of any one of embodiments 41-62, wherein the polynucleotide encoding the zika virus 2K or the derivative of the zika virus 2K comprises a sequence at least 80% identical to a 2K of Table 4, or a 2K from a zika virus of Table IB.

[0285] Embodiment 64. The zika virus-like particle or the composition of any one of embodiments 40 or 42-63, or the non-structural protein plasmid of any one of embodiments 41-63, wherein the polynucleotide encoding the derivative of the zika virus NS4B comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus NS4B.

[0286] Embodiment 65. The zika virus-like particle or the composition of any one of embodiments 40 or 42-64, or the non-structural protein plasmid of any one of embodiments 41-64, wherein the zika virus NS4B protein or the derivative of the zika virus NS4B comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus NS4B from Table 4, or a NS4B from a zika virus of Table IB.

[0287] Embodiment 66. The zika virus-like particle or the composition of any one of 40 or 42-65, or the non-structural protein plasmid of any one of embodiments 41-65, wherein the polynucleotide encoding the zika virus NS4B or the derivative of the zika virus NS4B comprisesa sequence at least 80% identical to a NS4B from Table 4, or a NS4B from a zika virus of Table IB.

[0288] Embodiment 67. The zika virus-like particle or the composition of any one of embodiments 40 or 42-66, or the non-structural protein plasmid of any one of embodiments 41-66, wherein the polynucleotide encoding the derivative of the zika virus NS5 comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus NS 5.

[0289] Embodiment 68. The zika virus-like particle or the composition of any one of embodiments 40 or 42-67, or the non-structural protein plasmid of any one of embodiments 41-67, wherein the zika virus NS5 or the derivative of the zika virus NS5 comprises an amino acid sequence at least 80% identical to an amino acid sequence encoded by a zika virus NS5 from Table 4, or a NS5 from a zika virus of Table IB.

[0290] Embodiment 69. The zika virus-like particle or the composition of any one of embodiments 40 or 42-68, or the non-structural protein plasmid of any one of embodiments 41-68, wherein the polynucleotide encoding the zika virus NS5 or the derivative of the zika virus NS5 comprises a sequence at least 80% identical to a NS5 from Table 4, or a NS5 from a zika virus of Table IB.

[0291] Embodiment 70. The zika virus-like particle or the composition of any one of embodiments 40 or 42-69, or the non-structural protein plasmid of any one of embodiments 41-69, wherein the second polynucleotide or the non-structural protein plasmid does not comprise a structural protein selected from (a) a zika virus capsid protein (C), or a derivative of a zika virus C (b) a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M (c) a zika virus viral envelope protein (E) or a derivative of a zika virus E, or (d) a combination of two or more selected from (a) to (c).

[0292] Embodiment 71. The zika virus-like particle or the composition of any one of embodiments 40 or 42-70, or the non-structural protein plasmid of any one of embodiments 41-70, wherein the second polynucleotide or the non-structural protein plasmid further comprises a structural protein selected from (a) a zika virus capsid protein (C), or a derivative of a zika virus C (b) a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M (c) a zika virus viral envelope protein (E) or a derivative of a zika virus E, or (d) a combination of two or more selected from (a) to (c).

[0293] Embodiment 72. The zika virus-like particle or the composition of embodiment 70, or the non-structural protein plasmid of embodiment 70, wherein the second polynucleotide or the non- structural protein plasmid does not comprise zika virus C, zika virus prM / M, or zika virus E.

[0294] Embodiment 73. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40 or embodiments 42-72, or the non- structural protein plasmid of any one of embodiments 41-72, wherein the non- structural protein is a non- structural protein from an African zika virus, Asian zika virus, or Brazilian zika virus, or a combination thereof.

[0295] Embodiment 74. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40 or embodiments 42-73, or the non- structural protein plasmid of any one of embodiments 41-73, wherein the non- structural protein is a non- structural protein from an African zika virus, Asian zika virus, or Brazilian zika virus, or a combination thereof.

[0296] Embodiment 75. The zika virus-like particle or the composition of any one of embodiments 73-74, or the non- structural protein plasmid of any one of embodiments 73-74, wherein the African zika virus is African MR766 strain.

[0297] Embodiment 76. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40 or embodiments 42-75, wherein the polynucleotide of interest is expressed from the third polynucleotide and the third polynucleotide is a polynucleotide of interest plasmid.

[0298] Embodiment 77. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40 or embodiments 42-76, comprising: a polynucleotide encoding a zika virus 5’ UTR or a derivative of the zika virus 5’ UTR, (b) a polynucleotide encoding a zika virus 3’ UTR or a derivative of the zika virus 3’ UTR, (c) a polynucleotide of interest, or (d) a combination of two or more selected from (a) to (c).

[0299] Embodiment 78. A polynucleotide of interest plasmid comprising: (a) a polynucleotide encoding a zika virus 5’ UTR or a derivative of the zika virus 5’ UTR, (b) a polynucleotide encoding a zika virus 3’ UTR or a derivative of the zika virus 3’ UTR, (c) a polynucleotide of interest, or (d) a combination of two or more selected from (a) to (c).

[0300] Embodiment 79. The zika virus-like particle or the composition of embodiment 77, or the polynucleotide of interest plasmid of embodiment 78, wherein the polynucleotide encoding a derivative of the zika virus 5’ UTR comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus 5’ UTR.

[0301] Embodiment 80. The zika virus-like particle or the composition of embodiment 77 or embodiment 79, or the polynucleotide of interest plasmid of embodiment 78 or embodiment 79, wherein the zika virus 5’ UTR or the derivative of the zika virus 5’ UTR comprises a sequence atleast 80% identical to a 5’ UTR sequence of Table 1A, Table IF, or a 5’ UTR of a zika virus of Table IB.

[0302] Embodiment 81. The zika virus-like particle or the composition of any one of embodiments 77 or 79-80, or the polynucleotide of interest plasmid of any one of embodiments 78-80, wherein the polynucleotide encoding the derivative of the zika virus 3’ UTR comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wildtype zika virus 3 ’ UTR.

[0303] Embodiment 82. The zika virus-like particle or the composition of any one of embodiments 77 or 79-81, or the polynucleotide of interest plasmid of any one of embodiments 78-81, wherein the zika virus 3’ UTR or the derivative of the zika virus 3’ UTR comprises a sequence at least 80% identical to a 3’ UTR sequence of Table 1A or Table IF, or a 3’ UTR of a zika virus of Table IB.

[0304] Embodiment 83. The zika virus-like particle or the composition of any one of embodiments 1-10, 12-40, 42-77, or 79-82, or the polynucleotide of interest plasmid of any one of embodiments 78-82, wherein the polynucleotide of interest encodes a therapeutic agent or a diagnostic agent.

[0305] Embodiment 84. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-83, or the polynucleotide of interest plasmid of any one of embodiments 78-83, wherein the therapeutic agent is an antibody -based therapeutic agent, a hormone, a cytokine, an inhibitor of an immune check point regulator, an antagonist of an immune checkpoint regulator, an immune stimulatory molecule, an agonist of an immune co-stimulatory molecule, or a peptide.

[0306] Embodiment 85. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-84, or the polynucleotide of interest plasmid of any one of embodiments 78-84, wherein the polynucleotide of interest encodes a gene editing tool.

[0307] Embodiment 86. The zika virus-like particle or the composition of embodiment 85, or the polynucleotide of interest plasmid of embodiment 85, wherein the gene editing tool is selected from a group comprising meganuclease associated agents, CRISPR associated agents, TALEN associated agents, and zinc finger associated agents.

[0308] Embodiment 87. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-84, or the polynucleotide of interest plasmid of any one of embodiments 78-84, wherein the polynucleotide of interest is a small interfering RNA (siRNA), antisense RNA, micro RNA (miRNA), small orshort hairpin RNA (shRNA), guide RNA (gRNA), clustered regularly interspaced short palindromic repeat RNA (crRNA), trans-activating clustered regularly interspaced short palindromic repeat RNA (tracrRNA), immune-stimulating oligonucleotide, antisense nucleic acid, or ribozyme.

[0309] Embodiment 88. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-84, or the polynucleotide of interest plasmid of any one of embodiments 78-84, wherein the polynucleotide of interest encodes a polypeptide associated with a genetic disorder.

[0310] Embodiment 89. The zika virus-like particle or the composition of embodiment 88, or the polynucleotide of interest plasmid of embodiment 88, wherein the polypeptide associated with a genetic disorder is brain-derived neurotrophin factor (BDNF), nerve growth factor (NGF), Neprilysin inhibitors (NEP), endothelin converting enzyme (ECE), cathepsin B (CTSB), apolipoprotein E 2 (APOE2), SH3 and multiple ankyrin repeat domains protein (SHANK), neurturin (NRTN), glial cell-derived neurotrophic factor (GDNF), cerebral dopamine neurotrophic factor (CDNF), vascular endothelial growth factor A (VEGF-A), or aromatic L- amino acid decarboxylase (AADC).

[0311] Embodiment 90. The zika virus-like particle or the composition of embodiment 84, or the polynucleotide of interest plasmid of embodiment 84, wherein the inhibitor of an immune checkpoint regulator or the antagonist of an immune checkpoint regulator is an anti-PDl antibody.

[0312] Embodiment 91. The zika virus-like particle or the composition of embodiment 84, or the polynucleotide of interest plasmid of embodiment 84, wherein the antibody -based therapeutic agent is an antibody, a functional fragment of an antibody, a chimeric antigen receptor (CAR), or a T cell receptor (TCR).

[0313] Embodiment 92. The zika virus-like particle or the composition of embodiment 84, or the polynucleotide of interest plasmid of 84, wherein the cytokine comprises lymphokine, monokine, polypeptide hormone, growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, glycoprotein hormone, follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), hepatic growth factor, fibroblast growth factor, prolactin, placental lactogen, tumor necrosis factor-a and -P, mullerian-inhibiting substance, mouse gonadotropin-associated peptide, inhibin, activin, vascular endothelial growth factor, integrin, thrombopoietin (TPO), nerve growth factor, NGF-P, platelet growth factor, transforming growth factor (TGF), TGF-a, TGF-P, insulin-like growth factor-1 and -II, erythropoietin (EPO), osteoinductive factor, interferon, such as interferon-a, -P and -y, colonystimulating factor (CSF), macrophage-CSF (M-CSF), granulocyte-macrophage-CSF (GM-CSF), granulocyte-CSF (GCSF), interleukin (IL), tumor necrosis factor, TNF-a, TNF-P, LIF, or kit ligand (KL), or a combination of two or more thereof.

[0314] Embodiment 93. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-84, or the polynucleotide of interest plasmid of any one of embodiments 78-84, wherein the polynucleotide of interest targets beta-secretase 1 (BACE1) and / or amyloid precursor protein (APP).

[0315] Embodiment 94. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-84, or the polynucleotide of interest plasmid of any one of embodiments 78-84, wherein the polynucleotide of interest encodes an antigen or an antigenic epitope thereof.

[0316] Embodiment 95. The zika virus-like particle or the composition of embodiment 94, or the polynucleotide of interest plasmid of embodiment 94, wherein the antigen or antigenic epitope thereof is disease associated.

[0317] Embodiment 96. The zika virus-like particle or the composition of embodiment 94 or embodiment 95, or the polynucleotide of interest plasmid of embodiment 94 or embodiment 95, wherein the antigen or antigenic epitope thereof is from a pathogen.

[0318] Embodiment 97. The zika virus-like particle or the composition of embodiment 96, or the polynucleotide of interest plasmid of embodiment 96, wherein the pathogen is a virus.

[0319] Embodiment 98. The zika virus-like particle or the composition of embodiment 97, or the polynucleotide of interest plasmid of embodiment 97, wherein the virus is a human SARS coronavirus, an influenza A virus, an influenza B virus, an influenza C virus an ebolavirus, a hepatitis B virus, a hepatitis C virus, a herpes simplex virus, a human immunodeficiency virus (HIV), a human papillomavirus (HPV-6, HPV-11), a measles virus, a rabies virus, a poliovirus, or a yellow fever virus.

[0320] Embodiment 99. The zika virus-like particle or the composition of embodiment 96, or the polynucleotide of interest plasmid of embodiment 96, wherein the pathogen is a bacterium.

[0321] Embodiment 100. The zika virus-like particle or the composition of embodiment 99, or the polynucleotide of interest plasmid of embodiment 99, wherein the bacterium is Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella. henselae, Bartonella quintana, Bifidobacterium Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacia, Burkholderia pseudomallei, Campylobacter jejuni, Cardiobacterium hominis, Campylobacter fetus, Chlamydia pneumonia, Chlamydia trachomatis, Clostridium difficile, Cyanobacteria, Eikenella corrodens, Enterobacter, Enterococcus faecium, Escherichiacoli, Escherichia coli 0157, Francesilla tularensis, Fusobacterium nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzae, Helicobacter pylori, Kingella kingae, Klebsiella pneumonia, Legionella bacteria, Legionella pneumophila serogroup 1, Leptospira, Morganella morganii, Neisseria gonorrhoeae, Neisseria meningitidis, Proteus mirabilis, Proteus vulgaris, Proteus myxofaciens, Providencia rettgeri, Providencia alcalifaciens, Providencia stuartii, Pseudomonas aeruginosa, Pseudomonas paucimobilis, Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas acidovorans, Rickettsiae, Salmonella enterica, Salmonella typhi, Salmonella paratyphi types A, B typhus, Salmonella, dublin, Salmonella arizonae, Salmonella choleraesuis, Serratia marcescens, Shigella dysenteriae, Shigella flexneri, Shigella boydii, Shigella sonnei, Treponema, Stenotrophomonas maltophilia, Vibrio cholerae, Vibrio mimicus, Vibrio alginolyticus, Vibrio hollisae, Vibrio parahaemolyticus, Vibrio vulnificus, Yersinia pestitis, Actinomycetes, Bacillus anthracis, Bacillus subtilis, Vibrio cholerae, Vibrio mimicus, Vibrio alginolyticus, Vibrio hollisae, Vibrio parahaemolyticus, Vibrio vulnificus, Yersinia pestitis, Actinomycetes, Bacillus anthracis, Bacillus subtilis, Mycoplasma, Nocardia, Propionibacterium, Pseudomonas aeruginosa, Pneumococci, Staphylococcus aureus, Staphylococcus epidermidis, methicillin resistant Staphylococcus aureus (MRSA), vancomycin resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcus saprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutants.

[0322] Embodiment 101. The zika virus-like particle or the composition of embodiment 96, or the polynucleotide of interest plasmid of embodiment 96, wherein the pathogen is a fungus, an amoeba, or a parasite.

[0323] Embodiment 102. The zika virus-like particle or the composition of embodiment 101, or the polynucleotide of interest plasmid of embodiment 101, wherein the fungus, amoeba, or parasite is Acanthamoeba spp, American trypanosomiasis, Balamuthia mandrillaris, Babesia divergens, Babesia bigemina, Babesia equi, Babesia microti, Babesia duncani, Balantidium coli, Blastocystis spp Cryptosporidium spp, Cyclospora cayetanensis, dientamoeba fragilis, Diphyllobothrium latum, Leishmania amazonesis, Naegleria fowderi, Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale curtisi, Plasmodium malariae, Rhinosporidium seeberi, Sarcocystis bovihominis, Sarcocystis suihominis, Toxoplasma gondii, Trichomonas vaginalis, Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0324] Embodiment 103. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-102, or thepolynucleotide of interest plasmid of any one of embodiments 78-102, wherein the polynucleotide of interest is about 5000 to 10000 nucleotides long.

[0325] Embodiment 104. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-103, or the polynucleotide of interest plasmid of any one of embodiments 78-103, wherein the polynucleotide of interest is about 5000 nucleotides long.

[0326] Embodiment 105. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-103, or the polynucleotide of interest plasmid of any one of embodiments 78-103, wherein the polynucleotide of interest is about 10000 nucleotides long.

[0327] Embodiment 106. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-105, or the polynucleotide of interest plasmid of any one of embodiments 78-105, wherein the polynucleotide of interest encodes a polypeptide comprising a polynucleotide sequence at least 80% identical to a sequence of Table IE.

[0328] Embodiment 107. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-106, or the polynucleotide of interest plasmid of any one of embodiments 78-106, wherein the polynucleotide of interest encodes a polypeptide selected from Table IE.

[0329] Embodiment 108. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-107, or the polynucleotide of interest plasmid of any one of embodiments 78-107, wherein the polynucleotide of interest comprises a polynucleotide sequence at least about 80% identical to a sequence of Table IE.

[0330] Embodiment 109. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-108, or the polynucleotide of interest plasmid of any one of embodiments 78-108, wherein the polynucleotide of interest plasmid comprises a polynucleotide encoding a zika virus C or a derivative of the zika virus C, wherein the derivative of the zika virus C comprises at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus C.

[0331] Embodiment 110. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-108, or the polynucleotide of interest plasmid of any one of embodiments 78-108, wherein thepolynucleotide of interest plasmid comprises a polynucleotide encoding a zika virus C or a derivative of the zika virus C, wherein the zika virus C or derivative of the zika virus C comprises an amino acid sequence encoding a sequence at least 80% identical to a sequence of Table 1 A, Table IF, or Table 2, or a zika virus C of a zika virus from Table IB.

[0332] Embodiment 111. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-108, or the polynucleotide of interest plasmid of any one of embodiments 78-108, wherein the polynucleotide of interest plasmid comprises a polynucleotide encoding a zika virus C or a derivative of the zika virus C, wherein the zika virus C or derivative of the zika virus C comprises a sequence at least 80% identical to a sequence of Table 1A, Table IF, or Table 2, or a zika virus C of a zika virus from Table IB.

[0333] Embodiment 112. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-111, or the polynucleotide of interest plasmid of any one of embodiments 78-111, wherein the polynucleotide of interest plasmid comprises a polynucleotide encoding a zika virus or a derivative of the zika virus prM / M comprising at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus prM / M.

[0334] Embodiment 113. The zika virus-like particle or the composition of embodiment 112, or the polynucleotide of interest plasmid of embodiment 112, wherein the zika virus prM / M protein or the derivative of the zika virus prM / M comprises an amino acid sequence at least 80% identical to a sequence of Table 1A, an amino acid sequence at least 80% identical to the amino acid sequence encoded by a prM of Table 3 or a prM of a zika virus of Table IB.

[0335] Embodiment 114. The zika virus-like particle or the composition of embodiment 112, or the polynucleotide of interest plasmid of embodiment 112, wherein the polynucleotide encoding the zika virus prM / M or the derivative of the zika virus prM / M comprises a sequence at least 80% identical to a sequence of Table 1 A or Table 3, or a prM / M of a zika virus from Table IB.

[0336] Embodiment 115. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-114, or the polynucleotide of interest plasmid of any one of embodiments 78-114, wherein the polynucleotide of interest plasmid comprises a polynucleotide encoding a zika virus E or a derivative of the zika virus E comprising at least one substitution, at least one deletion, and / or at least one insertion as compared to a wild-type zika virus E.

[0337] Embodiment 116. The zika virus-like particle or the composition of embodiment 115, or the polynucleotide of interest plasmid of embodiment 115, wherein the zika virus E protein or thederivative of the zika virus E comprises an amino acid sequence at least 80% identical to a sequence of Table 1A, an amino acid sequence at least 80% identical to the amino acid sequence encoded by a E of Table 3, or an E of a zika virus of Table IB.

[0338] Embodiment 117. The zika virus-like particle or the composition of embodiment 115, or the polynucleotide of interest plasmid of embodiment 115, wherein the polynucleotide encoding the zika virus E or the derivative of the zika virus E comprises a sequence at least 80% identical to a sequence of Table 1A or Table 3, or an E of a zika virus from Table IB.

[0339] Embodiment 118. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-117, or the polynucleotide of interest plasmid of any one of embodiments 78-117, wherein the polynucleotide of interest plasmid does not comprise a polynucleotide encoding one or more non- structural (NS) proteins selected from (i) a NS1, (ii) a NS2A, (iii) a NS2B, (iv) a NS3, (v) a NS4A, (vi) a NS4B, (vii) a NS 5, or (viii) two or more of (i)-(vii).

[0340] Embodiment 119. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-118, or the polynucleotide of interest plasmid of any one of embodiments 78-118, wherein the polynucleotide of interest plasmid further comprises a polynucleotide encoding one or more NS proteins selected from (i) a NSl, (ii) a NS2A, (iii) a NS2B, (iv) a NS3, (v) a NS4A, (vi) a NS4B, (vii) a NS5, or (viii) two or more of (i)-(viii).

[0341] Embodiment 120. The zika virus-like particle or the composition of embodiment 118 or embodiment 119, or the polynucleotide of interest plasmid of embodiment 118 or embodiment 119, wherein the NS1 is a zika virus NS1, the NS2A is a zika virus NS2A, the NS2B is a zika virus NS2B, the NS3 is a zika virus NS3, the NS4A is a zika virus NS4A, the NS4B is a zika virus NS4B, or the NS5 is a zika virus NS5.

[0342] Embodiment 121. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-120, or the polynucleotide of interest plasmid of any one of embodiments 78-120, wherein the structural protein is a structural protein from an African zika virus, Asian zika virus, or Brazilian zika virus, or a combination of two or more.

[0343] Embodiment 122. The zika virus-like particle or the composition of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-121, or the polynucleotide of interest plasmid of any one of embodiments 78-121, wherein the non- structural protein is a non-structural protein from an African zika virus, Asian zika virus, or Brazilian zika virus, or a combination of two or more.

[0344] Embodiment 123. The zika virus-like particle or the composition of embodiment 121 or embodiment 122, or the polynucleotide of interest plasmid of embodiment 121 or embodiment 122, wherein the African zika virus is African MR766 strain.

[0345] Embodiment 124. A method for producing a zika virus-like particle, comprising introducing into a host cell: (a) the structural protein plasmid of any one of embodiments 11-38, (b) the non- structural protein plasmid of any one of embodiments 41-75, and (c) the polynucleotide of interest plasmid of any one of embodiments 78-123.

[0346] Embodiment 125. A method for producing a zika virus-like particle, comprising introducing into a host cell the composition of any one of embodiments 1-10, 12-40, 42-77, or 79-123, wherein the first polynucleotide is a structural protein plasmid, the second polynucleotide is a non- structural protein plasmid, and the third polynucleotide is a polynucleotide of interest plasmid.

[0347] Embodiment 126. The method for producing the zika virus-like particle of embodiment 124 or embodiment 125, wherein the structural protein plasmid, non- structural protein plasmid, and polynucleotide of interest plasmid are introduced simultaneously.

[0348] Embodiment 127. The method for producing the zika virus-like particle of embodiment 124 or embodiment 125, wherein the structural protein plasmid, non- structural protein plasmid, and polynucleotide of interest plasmid are introduced in any order.

[0349] Embodiment 128. The method for producing the zika virus-like particle of any one of embodiments 124-127, wherein introducing into a host cell comprises use of a transfection reagent.

[0350] Embodiment 129. The method for producing the zika virus-like particle of any one of embodiments 124-128, wherein the transfection reagent is a lipid-based agent.

[0351] Embodiment 130. The method for producing the zika virus-like particle of embodiment 129, wherein the lipid-based agent is lipofectin, lipofectamine, cellfectin, polycations, liposomes, or a lipid nanoparticle.

[0352] Embodiment 131. The method for producing the zika virus-like particle of any one of embodiments 124-130, wherein the host cell is a mammalian cell.

[0353] Embodiment 132. The method for producing the zika virus-like particle of embodiment 131, wherein the mammalian cell is a kidney cell.

[0354] Embodiment 133. The method for producing the zika virus-like particle of embodiment 131, wherein the mammalian cell is a HEK cell line or VERO cell line.

[0355] Embodiment 134. The method for producing the zika virus-like particle of embodiment 131, wherein the mammalian cell is a Vero E6, HEK, HEK293T, HEK293TT, FreeStyle™ 293-F Cell, HEK-293.2sus or C6 / 36 cell.

[0356] Embodiment 135. The method for producing the zika virus-like particle of any one of embodiments 124-134, further comprising purifying the zika virus-like particle.

[0357] Embodiment 136. The method for producing the zika virus-like particle of embodiment 135, wherein purifying the zika virus-like particle comprises a sucrose cushion, tangential flow filtration, affinity chromatography, ion exchange chromatography, hydrophobic chromatography, size exclusion chromatography, or gel filtration, or a combination of two or more thereof.

[0358] Embodiment 137. The method for producing the zika virus-like particle of any one of embodiments 124-136, wherein the zika virus-like particles are precipitated with polyethylene glycol (PEG), trifluoroacetic acid, centrifugation, or ultracentrifugation.

[0359] Embodiment 138. A method of treating a disease or condition, comprising administering to a subject in need thereof the zika virus-like particle of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-123; or the zika virus-like particle produced by the method of any one of embodiments 124-137.

[0360] Embodiment 139. The method of treating a disease or condition of embodiment 138, wherein the zika virus-like particle has tropism for central nervous system, ocular system cell, or tumor cell.

[0361] Embodiment 140. The method of treating a disease or condition of embodiment 138 or embodiment 139, wherein administering to a subject in need thereof the zika virus-like particle comprises administering to a target cell.

[0362] Embodiment 141. The method of treating a disease or condition of embodiment 140, wherein the target cell is a neuronal cell.

[0363] Embodiment 142. The method of treating a disease or condition of embodiment 140, wherein the target cell is a non -neuronal cell.

[0364] Embodiment 143. The method of treating a disease or condition of embodiment 141, wherein the neuronal cell is an oligodendrocyte, microglia, or astrocyte.

[0365] Embodiment 144. The method of treating a disease or condition of embodiment 142, wherein the non-neuronal cell is a prostate epithelial cell, a urethra epithelial cells, a Sertoli cell, a Leydig cell, a spermatogonium cell, or a retinal cell.

[0366] Embodiment 145. The method of treating a disease or condition of any one of embodiments 138-144, wherein the disease or condition is a disease or condition of the brain.

[0367] Embodiment 146. The method of treating a disease or condition of any one of embodiments 138-144, wherein the disease or condition is a disease or condition of the ocular system.

[0368] Embodiment 147. The method of treating a disease or condition of any one of embodiments 138-145, wherein the disease or condition is Alzheimer’s disease.

[0369] Embodiment 148. The method of treating a disease or condition of any one of embodiments 138-144, wherein the disease or condition is metabolic dysfunction.

[0370] Embodiment 149. The method of treating a disease or condition of any one of embodiments 138-145, wherein the disease or condition is Parkinson’s disease.

[0371] Embodiment 150. The method of treating a disease or condition of any one of embodiments 138-145, wherein the disease or condition is autism.

[0372] Embodiment 151. The method of treating a disease or condition of any one of embodiments 138-145, wherein the disease or condition is Down’s Syndrome.

[0373] Embodiment 152. The method of treating a disease or condition of any one of embodiments 138-146, wherein the disease or condition is choroideremia-blindness.

[0374] Embodiment 153. The method of treating a disease or condition of any one of embodiments 138-146, wherein the disease or condition is leber congenital amaurosis.

[0375] Embodiment 154. The method of treating a disease or condition of any one of embodiments 138-146, wherein the disease or condition is ornithine transcarbamylase (OTC) deficiency.

[0376] Embodiment 155. The method of treating a disease or condition of any one of embodiments 138-145, wherein the disease or condition is multiple sclerosis.

[0377] Embodiment 156. The method of treating a disease or condition of any one of embodiments 138-144, wherein the disease or condition is Pompe Disease.

[0378] Embodiment 157. The method of treating a disease or condition of any one of embodiments 138-145, wherein the disease or condition is depression.

[0379] Embodiment 158. The method of treating a disease or condition of any one of embodiments 140-157, wherein the target cell transiently expresses the polynucleotide of interest after delivery.

[0380] Embodiment 159. The method of treating a disease or condition of any one of embodiments 140-157, wherein the target cell persistently expresses the polynucleotide of interest after delivery.

[0381] Embodiment 160. The method of treating a disease or condition of any one of embodiments 138-159, wherein the method comprises administering to the subject an effectiveamount of the zika virus-like particle comprising a polynucleotide of interest encoding a brain derived neurotrophin factor (BDNF).

[0382] Embodiment 161. The method of treating a disease or condition of any one of embodiments 138-159, wherein the method comprises administering to the subject an effective amount of the zika virus-like particle comprising an polynucleotide of interest encoding a brain derived neurotrophin factor (BDNF).

[0383] Embodiment 162. The method of treating a disease or condition of any one of embodiments 138-159, wherein the method comprises administering to the subject an effective amount of the zika virus-like particle comprising a polynucleotide of interest encoding a SH3 and multiple ankyrin repeat domains protein (SHANK).

[0384] Embodiment 163. The method of treating a disease or condition of any one of embodiments 138-159, wherein the method comprises administering to the subject an effective amount of the zika virus-like particle comprising a polynucleotide of interest encoding an aromatic L-amino acid decarboxylase (AADC).

[0385] Embodiment 164. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof is via systemic delivery.

[0386] Embodiment 165. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof is performed intravenously.

[0387] Embodiment 166. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof is performed intratumorally.

[0388] Embodiment 167. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof is performed intravenously and intratumorally.

[0389] Embodiment 168. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof targets the cerebral spinal fluid.

[0390] Embodiment 169. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof is performed via intracerebroventri cul ar admini strati on .

[0391] Embodiment 170. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof is performed via cistema magna administration.

[0392] Embodiment 171. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof is performed via subpial administration.

[0393] Embodiment 172. The method of treating a disease or condition of any one of embodiments 138-163, wherein the administering to a subject in need thereof is performed via intrathecal administration.

[0394] Embodiment 173. The method of treating a disease or condition of any one of embodiments 138-172, wherein the administering to a subject in need thereof is not performed via intraparenchymal administration.

[0395] Embodiment 174. A method of triggering or boosting an immune response, the method comprising administering to a subject in need thereof an effective amount of the zika virus-like particle of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-123; or the zika virus-like particle produced by the method of any one of embodiments 124-137.

[0396] Embodiment 175. The method of triggering or boosting an immune response of embodiment 174, wherein the polynucleotide of interest comprises an antigen or an antigenic epitope thereof.

[0397] Embodiment 176. The zika virus-like particle of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, or embodiments 79-123, or the zika virus-like particle produced by the method of any one of embodiments 124-137, wherein the zika virus-like particle has tropism for tumor cells.

[0398] Embodiment 177. The zika virus-like particle of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, embodiments 79-123, or the zika virus-like particle produced by the method of any one of embodiments 124-137, wherein the zika virus-like particle has tropism for central nervous system or ocular system cells.

[0399] Embodiment 178. A vaccine composition comprising the zika virus-like particle of any one of embodiments 1-10, embodiments 12-40, embodiments 42-77, embodiments 79-123, or the zika virus-like particle produced by the method of any one of embodiments 124-137.

[0400] Embodiment 179. The method of treating a disease or condition of any one of embodiments 138-173, wherein the disease or condition is cortex and medulla disorders.

[0401] Embodiment 180. The method of treating a disease or condition of any one of embodiments 138-173, wherein the disease or condition is single-gene inheritance disease.

[0402] Embodiment 181. The method of treating a disease or condition of any one of embodiments 138-173, wherein the disease or condition is multifactorial genetic inheritance disease.EXAMPLESExample 1: Evaluation of Zika capsid coding sequence

[0403] The evaluation of Zika capsid sequence necessary for encapsulation of a polynucleotide comprising a polynucleotide of interest (e.g., pREP) was carried out with five different plasmid constructs (FIGS. 2A-2E). Each plasmid comprises a Eukaryotic promoter (e.g., CMV promoter), RNA polymerase promoter (e.g., T7 promoter), first ribozyme (e.g., Hammerhead ribozyme), 5’ UTR, a sequence encoding for part of a zika virus C protein (C-partial (15, 30, 45, 60 or 75 bp)) or without C-partial, 3’ UTR, second ribozyme (e.g., HDV ribozyme), and sequences encoding reporter proteins (e.g., GFP and Nluc luciferase). See Table IF and Table 2 below for the C- partial and plasmid component sequences.

[0404] Using a kit MEGAscript (catalog #AM1330, Life Technologies), in vitro transcription was performed for the vectors according to the user manuals.

[0405] Vero cells were obtained from ATCC and were raised under DMEM high culture medium. After reaching 80% confluency, RNA (FIGS. 2A-2E, or a plasmid not coding a capsid) was transfected. The RNA transfection was carried out with Lipofectamine™ MessengerMAX™ (Thermo Scientific™ catalog # LMRNA001) under fabricant instruction conditions.

[0406] After 24 hours, Vero cells were infected with the wild type zika virus. The Vero cells were infected with a virus concentration of MOI 0.01. After 30 minutes of the incubation period, the culture infection buffer was replaced with fresh media.

[0407] After 120 hours after ZIKV infection, the culture supernatant, containing the viral vectors (ZIKVV) produced by Vero cells, was collected.

[0408] RNA quantification was performed from different viral vectors generated (FIGS. 2A- 2E, or a plasmid not coding a capsid).

[0409] RT-PCR was performed with specific primers for the ZIKV genome and pREP (pREP gene) to detect the RNA amount of the ZIKV genome and pREP after pREP transfection and ZIKV infection in Vero cells. pREP RNA copies were quantified from all pREP with differentsized capsid sequences (FIG. 3). FIG. 3 shows pREP RNA encapsulation in all plasmids tested (C-partial with no Capsid sequence, 15, 30, 45, 60 and 75 bp). The pREP RNA encapsulationwas higher in C-partial with 15, 45, 60 and 75 bp, indicating that ZIKVV carrying pREP RNA with different capsid partial length change the RNA efficiency encapsulated into the viral vector.

[0410] To test the viral vector delivery of pREP with different size capsid sequences into a target cell, the supernatant was contacted with Vero cell lines to confirm Nanoluc delivery by ZIKVV. The graph in FIG. 4 shows Nanoluc quantification in Vero cells after contact with viral vector (pREP + ZIKV) carrying no C-partial sequence, 5, 10, 15, 20 and 25 amino acids, derived from RNA (FIG. 3) transfection, compared to ZIKV (only ZIKV infection). A higher level of NanoLuc expression is demonstrated in viral vectors of all capsid sequence sizes compared to the control group. These results indicate that ZIKVV is able to encapsulate pREP RNA with C- partial (no C-partial sequence, 15, 30, 45, 60 and 75 bp) and delivery NanoLuc, confirmed by the expression of the reporter gene in the target cells.Table IF. Figure 1 sequences.Table 2: Zika capsid sequence for pREP (FIGS. 2A-2E) encapsulation.Example 2: DNA transfection of a construct carrying structural proteins and a construct carrying non-structural proteins in Vero and HEK-293 cells.

[0411] A first plasmid construct comprising a DNA polymerase promoter (e.g., CMV promoter), RNA polymerase promoter (e.g., T7 promoter), 5’ UTR (e.g., Xenopus globin 5’ UTR), internal ribosome entry site (IRES), 3’ UTR, and encoding Capsid, Membrane, Envelope and Nluc luciferase was designed as shown generally in FIG. 5. See also Table 3. In an example, this first plasmid is a structural protein plasmid.

[0412] A second plasmid construct comprising DNA polymerase promoter (e.g., CMV promoter), RNA polymerase promoter (e.g., T7 promoter), 5’ UTR, 3’ UTR, and encoding Nluc luciferase, Envelope-partial protein (E), non-structural protein 1 (NS1), non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, non-structural protein 2B (NS2B), non- structural protein 3 (NS3), non-structural protein 4 A (NS4A), non-structural protein 2K (2K), non-structural protein NS4B (NS4B) and non-structural protein NS5 (NS5) was designed as shown generally in FIG. 6. See also Table 4. In an example, this second plasmid is a non- structural protein plasmid, or a helper plasmid.

[0413] A third plasmid construct comprising a polynucleotide encoding a zika virus 5’ UTR or a derivative of the zika virus 5’ UTR, a polynucleotide encoding a zika virus 3’ UTR or a derivative of the zika virus 3’ UTR, a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C, a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M, a polynucleotide encoding a zika virus viral envelope protein E or a derivative of a zika virus E, a polynucleotide of interest, a eukaryotic enhancer, a eukaryotic promoter, one or more multiple cloning sites (MCS), a hammerhead, a ribozyme, and a poly(A) signal, as shown in FIGS 2A-2E.

[0414] In an example, this third plasmid is a pREP plasmid.

[0415] DNA from each construct was transfected in Vero and HEK-293 cells. The transient transfection was carried out with Lipofectamine™ MessengerMAX™ (Thermo Scientific™ catalog # LMRNA001).

[0416] After 24-, 48- and 72-hours Transfected cells were resuspended in 1 volume of Nano- Glo® Luciferase Assay System (Promega catalog # N1110) previously diluted lOx in water. Luminescence was quantified for 0.3 seconds in a GloMax luminometer (Promega). As shown in FIG. 7A and FIG. 7B, high Nluc activities were observed in both the cell types (Vero and HEK- 293 cells) and in all transfected plasmids (FIGS. 1, 5 and 6) demonstrating that the plasmids can express a reporter gene, thus validating the plasmids' functionality.

[0417] As shown in FIG. 7A and FIG. 7B, Nluc activities were higher in HEK-293 cells than in Vero cells. These results indicate that HEK-293 cells can express higher number of structural proteins and pREP RNA necessary for virus-like particle assemble. Non- structural plasmids (FIG. 6, Table 4) are more lengthy than Structural proteins (FIG. 5, Table 3) and pREP plasmids (FIG. 1, Table 1), which explains why Nluc activity is lower when compared to others.Table 3. Figure 5 sequences.Table 4. Figure 6 sequences.Example 3: Protein expression validation of structural and non-structural proteins for viruslike particles.

[0418] A transfection of Vero and HEK cells, or other eukaryotic cells is performed with a first plasmid containing structural protein coding sequence(s) and a second plasmid containing non- structural protein coding sequence(s).

[0419] A transfection of Vero and HEK cells, or other eukaryotic cells is performed with a plasmid containing structural protein coding sequence(s).

[0420] After 24-, 72- and 120-hours post DNA transfection, samples from cell culture medium and cell lysate are collected and the total protein sample is analyzed by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and the protein migration pattern is observed after Coomassie blue protein staining. The presence of the NS1-NS5 in the electrophoresis gel, shows the correct expression of the proteins for helping the production of the zika virus-like particles.

[0421] After 24-, 72- and 120-hours post DNA transfection, samples from cell culture medium and cell lysate are collected and the total protein sample is analyzed by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and the protein migration pattern is observedafter Coomassie blue protein staining. The presence of the, approximately 11.6 kDa, 19 kDa and 54.4 kDa protein from Capsid protein (C), Membrane protein (M) and Envelope protein (E), respectively, in the electrophoresis gel, shows the correct expression of the protein(s) for the production of the zika-virus-like particles.

[0422] Western blot analysis, using primary antibodies against C, M, and E proteins shows the expression of the recombinant capsid proteins for the correct formation of the Zika virus-like particles.Example 4: Production of zika virus-like particles.

[0423] A co-transfection of Vero and HEK cells, or other eukaryotic cells with three plasmids (structural, non- structural and pREP plasmids) is performed. Co-transfection is performed in groups with the plasmids alone and any combination of two plasmids to verify and guarantee that zika virus-like particles will encapsulate just the pREP RNA. As the pREP RNA will be the only one that will provide a cis-packing signal as opposed to other RNA, it will be encapsulated, therefore producing virus-like particles loaded selectively with the desired RNA.

[0424] In one experiment, the polypeptide of interest plasmid will be a plasmid as shown in FIGS. 1, 2A-2E, wherein the polypeptide of interest will be a reporter. The reporter will be EGFP and Nanoluciferase.

[0425] In one experiment, the structural protein plasmid will be a plasmid as shown in FIG. 5, wherein one of the proteins expressed will be a reporter. The reporter will be Nanoluciferase.

[0426] In another experiment, the structural protein plasmid will be a plasmid as shown in FIG.8, wherein one of the proteins expressed will not be a reporter.

[0427] In one experiment, the non-structural protein plasmid will be a plasmid as shown in FIG.6. In one experiment, the non-structural protein plasmid will be a plasmid as shown in FIG. 6, without a gene encoding a reporter.

[0428] After 24-, 72- and 120-hours post DNA transfection, The supernatant is purified with sucrose cushion and precipitated with PEG.

[0429] Samples from cell culture medium and cell lysate are collected and the total protein sample is separated by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS- PAGE) and the protein migration pattern is observed after Coomassie blue protein staining.

[0430] Western blot analysis is performed using primary antibodies against structural and non- structural proteins.

[0431] An aliquot of supernatant purified is subject to RNA extraction to carry out RT-PCR to detect quantitively the amount of pREP that is encapsulated in the viral vector productionprocess. Therefore, TaqMan One-Step Real-Time RT-PCR analysis is carried out that will be able to detect with high-sensitivity, even very low amounts of pREP RNA. All the methodology described above will be compared with the strategy that generates viral vectors from ZIKV wild type.

[0432] To verify if the virus-like particle can deliver the pREP RNA, the vector is incubated with Vero cells to observe the delivery and expression of NanoLuc enzyme. A lysate is used to analyse the amount of pREP RNA delivered in the target cell.Example 5: Structural analysis of the Zika virus-like particles by Transmission Electron Microscopy (TEM).

[0433] After the transfection with the structural protein plasmid and non-structural protein plasmid in Vero and HEK cells, the supernatant cell culture medium is collected and analyzed by TEM. The VLPs should exhibit particles size from 50nm to 65nm and morphology similar to the wild-type Zika virus.Example 6: Evaluation of the Zika virus-like in vitro tropism for central nervous cells (CNS).

[0434] A method is performed to evaluation whether the tropism of the viral vector (zika viruslike particle) persists in cells characterized by tropism for the ZIKA virus. The tests are carried out in cell lines DU-145, DAOY, CHLA, MC-F7, MDA-MB-231 and HCT8. DU-145, DAOY, CHLA can bind with the wild-type ZIKV, on the other hand, MC-F7, MDA-MB-231 and HCT8 lines, do not have the ability to bind the wild virus.

[0435] If the viral vector retains the ability to enter cell lines as does wild-type ZIKV, the RNA with reporter gene that codes the enzyme NanoLuc Luciferase will be delivered and translated, being detected through luminescence assay. In addition to luminescence analysis, TaqMan One- Step Real-Time RT-PCR analysis is carried out to detect the presence of pREP RNA after contact with the viral vector, even in very low amounts of viral nucleic acid or pREP RNA.Example 7: In vivo validation of zika virus-like particle of reporter gene expression and tropism to the brain.

[0436] To confirm the in vivo tropism of the zika virus-like particle, Balb / C Nude mice, an immunosuppressed animal model, without a thymus, susceptible to ZIKV infection, are tested.

[0437] Since these mice models have confirmed ZIKV susceptibility and SC tropism, we compare the in vivo behavior of zika virus-like particle. Each treatment group is randomly assigned to a cage so that different groups / treatments are exposed to the same environment andany batch effects are minimized. In these studies, the experimental unit for analysis is the mouse and the viral vectors injections with each respective control are: ZIKVV generated from ZIKV wild-type cell infection (ZIKVV-WT) versus negative control (Brazilian ZIKV stream - ZIKV) in comparison with zika virus-like particle, with and without pREP (experimental group: ZIKV; ZIKVV / WT; VLP / pREP+; VLP / pREP-). Following injection, animals are monitored for one hour for potential adverse events, including lethargy, respiratory changes, or other abnormal behavior. 24 hours after injection, the mice brain is collected for histopathological analysis of P- Galactosidase (P-gal) by treating the paraffinized histological sections with P-gal substrate. The positive staining of P-gal in the brain tissue indicates tropism of the virus or viral-like particle to the brain.Example 8: Evaluation of Zika capsid coding sequence

[0438] The evaluation of Zika capsid sequence necessary for encapsulation of a polynucleotide comprising a polynucleotide of interest (e.g., pREP) is carried out with six different plasmid constructs (FIGS. 10A-10F). Each plasmid comprises a Eukaryotic promoter (e.g., CMV promoter), RNA polymerase promoter (e.g., T7 promoter), first ribozyme (e.g., Hammerhead ribozyme), 5’ UTR, a sequence encoding for part of a zika virus C protein (C-partial (15, 30, 45, 60 or 75 bp)) or without C-partial, a sequence encoding for part of a zika virus NS4a-2K-NS4b protein or without NS4a2KNS4b-partial, 3’ UTR, second ribozyme (e.g., HDV ribozyme), and sequences encoding reporter proteins (e.g., GFP and Nanoluciferase). See Table 5, and Table 6 below for the C-partial and plasmid component sequences.

[0439] Using a kit MEGAscript (catalog #AM1330, Life Technologies), in vitro transcription is performed for the vectors according to the protocol.

[0440] Vero cells after reaching 80% confluency, RNA (FIGS. 10A-10F) is transfected. The RNA transfection is performed with Lipofectamine™ MessengerMAX™ (Thermo Scientific™ catalog # LMRNA001) under fabricant instruction conditions.

[0441] After 24 hours, Vero cells are infected with the wild-type Zika virus. The Vero cells were infected with a virus concentration of MOI 0.01. After 30 minutes of incubation, the culture infection buffer is replaced with fresh media.

[0442] 120 hours after ZIKV infection, the culture supernatant, containing the viral vectors (ZIKVV) produced by Vero cells, is collected.

[0443] RT-PCR is performed with specific primers for the ZIKV genome and pREP (pREP gene) to detect the RNA amount of the ZIKV genome and pREP after pREP transfection and- I l l -ZIKV infection in Vero cells. pREP RNA copies are quantified from all pREP with differentsized capsid sequences.

[0444] To test the viral vector delivery of pREP with different size capsid sequences into a target cell, the supernatant is contacted with Vero cell lines to confirm Nanoluciferase delivery by ZIKVV. Nanoluciferase is quantified in Vero cells after contact with viral vector (pREP + ZIKV) carrying no C-partial sequence, 5, 10, 15, 20, and 25 amino acids, derived from RNA transfection, compared to ZIKV (only ZIKV infection).Table 5. FIG. 9 sequences.Table 6: Zika capsid sequence for pREP (FIGS. 10A-10F) encapsulation.Table 9: Zika polypeptide sequence for full-length and partial non-structural protein of FIG. 9Example 9: DNA transfection of a construct carrying structural proteins and a construct carrying non-structural proteins in Vero and HEK-293 cells.

[0445] A first plasmid construct comprising a DNA polymerase promoter (e.g., CMV promoter), RNA polymerase promoter (e.g., T7 promoter), Xenopus globin 5’ UTR, internal ribosome entry site (IRES), Xenopus globin 3’ UTR, bGH poly(A) signal, and encoding Capsid, Membrane, Envelope and Nluc luciferase was designed as shown generally in FIG. 11. See also Table 7. In an example, this first plasmid is a structural protein plasmid.

[0446] A second plasmid construct comprising DNA polymerase promoter (e.g., CMV promoter), RNA polymerase promoter (e.g., T7 promoter), TEV 5’ UTR, Xenopus globin 3’ UTR, bGH poly(A) signal, and encoding Nluc luciferase, Envelope-partial protein (E), non- structural protein 1 (NS1), non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A, non-structural protein 2B (NS2B), non-structural protein 3 (NS3), non-structural protein4 A (NS4A), non-structural protein 2K (2K), non- structural protein NS4B (NS4B) and non- structural protein NS5 (NS5) was designed as shown generally in FIG. 12. See also Table 8. In an example, this second plasmid is a non-structural protein plasmid, or a helper plasmid.

[0447] A third plasmid is a pREP plasmid, as shown in FIG. 9.

[0448] DNA from each construct was transfected in Vero and HEK-293 cells. The transient transfection was performed with Lipofectamine™ MessengerMAX™ (Thermo Scientific™ catalog # LMRNA001).

[0449] After 24-, 48- and 72-hours Transfected cells were resuspended in 1 volume of Nano- Glo® Luciferase Assay System (Promega catalog # N1110) previously diluted lOx in water.Luminescence was quantified for 0.3 seconds in a GloMax luminometer (Promega). As shown in FIG. 7A and FIG. 7B, high Nanoluciferase activities were observed in both the cell types (Vero and HEK-293 cells) and in all transfected plasmids (FIGS. 9, 11, and 12) demonstrating that the plasmids can express a reporter gene, thus validating the plasmids' functionality.

[0450] As shown in FIG. 7A and FIG. 7B, Nanoluciferase activities were higher in HEK-293 cells than in Vero cells. These results indicate that HEK-293 cells can express higher number of structural proteins and pREP RNA necessary for virus-like particle assembly. Non-structural plasmids (FIG. 12, Table 8) are more lengthy than Structural proteins (FIG. 11, Table 7) and pREP plasmids (FIG. 9, Table 5), which explains why Nanoluciferase activity is lower when compared to others.Table 7. FIG. 11 sequences.Table 8. FIG. 12 sequences.Example 10: Production of Zika virus-like particles (ZIKW)

[0451] Co-transfection of Vero and HEK 293T cells, with three plasmids (structural, non- structural, and pREP plasmids) was performed as shown in FIG. 13B. In one experiment, the polypeptide of interest was encoded by a plasmid as shown in FIG. 9, where the polypeptide of interest was a reporter. The reporters used were EGFP and Nanoluciferase. The structural protein plasmid, as shown in FIG. 11, expressed a reporter protein, Nanoluciferase. The non- structural protein plasmid was as depicted in FIG. 12.

[0452] A mixture of plasmids consisting of DNA from each construct was transfected into Vero and HEK-293T cells using the Lipofectamine 3000 reagent (Thermo Scientific™, catalog # L3000001). The transfection solution containing the plasmids was mixed with the cells in DMEM supplemented with fetal bovine serum. Supernatants were collected at 24-, 48-, and 72- hours post-transfection.

[0453] In another experiment, a plasmid mixture consisting of DNA from each construct was transfected in HEK-293T cells and after 72- and 96-hours post-DNA transfection, The supernatant was collected (FIG. 16).

[0454] The supernatants were used both for RT-qPCR analysis and for incubation in Vero cells to verify the delivery of the reporter gene. For RT-qPCR, RNA was extracted from the supernatants using the QIAamp Viral RNA Mini Kit, following the manufacturer’s protocol. RNA copies were quantified by RT-qPCR using the One Step Real-Time PCR System and TaqMan™ Fast Virus 1-Step Master Mix. (FIG. 14A and FIG. 14B). Additionally, the number of copies detected for each plasmid was higher in HEK-293T cells compared to Vero cells.

[0455] To verify if the virus-like particle can deliver the pREP RNA, the vector was incubated with HEK-293T (FIG. 15B) and Vero cells (FIG. 15A) to observe the delivery and expression of the Nanoluciferase enzyme. It was confirmed the delivery of the pRep RNA containing the gene of interest by the viral vector (sZIKV-V). This was demonstrated by the quantification of bioluminescence, which was higher in the group treated with the modified viral vector compared to the negative control group, where the supernatant containing only pRep was applied to Vero cells. Additionally, it was observed that the viral vector collected 96 hours post-transfection exhibited better transduction efficiency compared to the 72-hour post-transfection collection (FIG. 16)Example 11: Protein expression validation of structural and non-structural proteins for viruslike particles

[0456] Co-transfection of HEK 293T cells, or other eukaryotic cells, with three plasmids (structural, non-structural, and pREP plasmids) was performed as shown in FIG. 13B. In this experiment, the polypeptide of interest was encoded by a plasmid as shown in FIG. 9, where the polypeptide of interest was a reporter. The reporters used were GFP and Nluc. The structural protein plasmid, as shown in FIG. 11, expressed a reporter protein, Nluc. The non-structural protein plasmid was as depicted in FIG. 12.

[0457] A mixture of plasmids consisting of DNA from each construct was transfected into Vero and HEK-293T cells using the Lipofectamine 3000 reagent (Thermo Scientific™, catalog # L3000001). The transfection solution containing the plasmids was mixed with the cells in DMEM supplemented with fetal bovine serum. Supernatants were collected at 96-hours posttransfection, The supernatant was collected and lysate.

[0458] The supernatant cell lysate was collected, and the total protein sample was analysed using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). The protein migration patterns were then visualized following Coomassie blue staining.

[0459] The detection of an approximately 54.4 kDa protein, identified as the Envelope protein (E), on the electrophoresis gel confirms the proper expression of the proteins necessary for generating Zika virus-like particles. Western blot analysis, using primary antibodies against Envelope protein shows the expression of the recombinant envelope protein for the correct formation of the Zika virus-like particles (FIG. 17).Example 12: Evaluation of the Zika virus-like particle delivery in vitro.

[0460] Three cell lines were employed in this experiment: MCF7, Vero, and CHLA-06. The MCF7 cell line is not susceptible to wildtype zika virus (ZIKV). The Vero cell is susceptible to wildtype zika virus and permits complete viral replication. Lastly, the CHLA-06 line, isolated from the cerebellum of a patient with Atypical Teratoid / Rhabdoid Tumor, is also susceptible and allows the replication of wildtype zika virus.

[0461] A mixture of plasmids consisting of DNA from each construct was transfected into Vero and HEK-293T cells using the Lipofectamine 3000 reagent (Thermo Scientific™, catalog # L3000001). The transfection solution containing the plasmids was mixed with the cells in DMEM supplemented with fetal bovine serum. Supernatants were collected at 72-hours posttransfection.

[0462] The viral vector was introduced to the cells and incubated at 37°C in 5% of CO2. This controlled environment facilitates the initial contact and subsequent entry of the viral vector into the cells, optimizing the conditions for the successful delivery of the vector's genetic material.

[0463] The assay results involving the MCF7 cell line once again highlighted a significant disparity in the modes of activity between the wild virus and the viral vector. Since the viral vector (sZIKV-V) is non-replicative, only the entry and delivery of its RNA are relevant to its mechanism of action. In contrast, the wild virus requires a conducive intracellular environment to complete its viral cycle, even though it can enter the cell. These results showed expands the potential target cell spectrum for the viral vector platform as compared to using a wildtype zika virus as a gene delivery modality (FIG. 18).Example 13: In vivo validation of zika virus-like particle of reporter gene expression and tropism to the brain

[0464] To confirm the in vivo tropism of the zika virus-like particle, BALB / c mice, an immunocompetent animal model, are tested. Since these mice models have confirmed ZIKV susceptibility and central nervous system tropism, we compare the in vivo behavior of zika viruslike particle. Each treatment group is randomly assigned to a cage so that different groups / treatments are exposed to the same environment and any batch effects are minimized.

[0465] In these studies, the experimental unit for analysis is the mouse and the viral vectors injections with each respective control are: zika virus-like particle formed from pREP, pStructural, and pNSP and wild type Zika virus. Following injection, animals are monitored for one hour for potential adverse events, including lethargy, respiratory changes, or other abnormal behavior.

[0466] At each collection time point, the mice were euthanized by anesthetic overdose. From each animal, blood and brain were collected. The blood was allowed to clot at room temperature for 30 minutes. The clot was then removed by centrifuging at 2,000 x g for 10 minutes at 4°C. The serum samples were used to analyze 48 protein targets in a single well using the ProcartaPlex™ Mouse Immune Monitoring Panel, 48plex (Thermo), and were analyzed with the Bio-Plex® MAGPIX™ Multiplex Reader (FIG. 19).

[0467] Mice that received treatments with viral vectors exhibited reduced profiles of pro- inflammatory cytokine expression compared to those treated with the wild-type virus. Specifically, the group treated with the wild-type virus showed higher expression of cytokines such as INF-alpha, IL-1 beta, IL-6, IL-7, IL-9, IL- 12, IL-22, and IL-33, which are associated with the innate response to natural viral infection (FIG. 19).

[0468] Furthermore, it was observed that the viral vector elicited a reduced immune response against it 14 days post-injection, unlike the persistent protein expression profile triggered by the wild-type virus (FIG. 19).

[0469] In another experiment the brain was collected and used to prepare histological slides and treated for immunofluorescence assay using DAPI for nuclear staining, anti-GFP antibody (sc- 996, Santa Cruz) for detection of GFP delivery and expression by pREP, and anti-NS2 (GTX133308, Genetex) for detection of ZIKV NS2 protein. For secondary labeling, AlexaFluor (AF)-conjugated antibodies were used, such as AF488 (Al 1008, Thermo), to mark NS2, and AF647 (A21235, Thermo) to mark GFP. Immunofluorescence images are obtained using a confocal microscope. Through immunofluorescence analysis, it was possible to verify the ability of sZIKV-V viral vector to cross the blood-brain barrier (BBB) and deliver pREP to brain cells in the hippocampus and cortex region.

[0470] The presence of the red marking shows the expression of GFP in both viral vectors within 3, 7, and 14 days, confirming the delivery of the gene of interest (GFP), while the green marking is present in animals that received ZIKV, as expected since it carries the genetic information necessary for the expression of NS2. The lack of detection of NS2 at sZIKV-V group confirms the correct encapsulation of only pREP and not for the NSPs RNA construct (FIG.20). The preceding merely illustrates the principles of this disclosure. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of this disclosure and the concepts contributed by the inventors to furthering the art, and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. The scope of the present disclosure, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of the present disclosure is embodied by the appended claims.

Claims

CLAIMSWhat is claimed is:

1. A zika virus-like particle, comprising: a polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the polynucleotide does not encode one or more viral non- structural proteins and one or more viral structural proteins.

2. The zika virus-like particle of claim 1, wherein the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, and zika virus NS5 or a derivative of the zika virus NS 5; and wherein the one or more viral structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, and zika virus viral envelope protein (E) or a derivative of the zika virus E.

3. A method of producing the zika virus-like particle of claim 1 or claim 2, the method comprising introducing into a host cell: the polynucleotide of claim 1 or claim 2, a second polynucleotide encoding the one or more viral non-structural proteins, and a third polynucleotide encoding the one or more viral structural proteins.

4. The method of claim 3, wherein each of the second polynucleotide and the third polynucleotide independently encode (i) a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus, and (ii) a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus.

5. The zika virus-like particle of claim 1, wherein the one or more viral non-structural proteins comprises zika virus NS1 or a derivative of the zika virus NS1, zika virus NS2a or a derivative of the zika virus NS2a, zika virus NS2b or a derivative of the zika virus NS2b, zika virus NS3 or a derivative of the zika virus NS3, or zika virus NS5 or a derivative of the zika virus NS5, or a combination of two or more thereof.

6. The zika virus-like particle of claim 1 or claim 5, wherein the polynucleotide encodes a zika virus NS4a comprising a sequence at least 90% identical to SEQ ID NO: 2929 or 2946.

7. The zika virus-like particle of any one of claims 1, 5, or 6, wherein the polynucleotide encodes a zika virus NS4a comprising a pTMS3.

8. The zika virus-like particle of any one of claims 1 or 5-7, wherein the polynucleotide does not encode a zika virus NS4a comprising a pTMSl and a pTMS2.

9. The zika virus-like particle of any one of claims 1 or 5-8, wherein the polynucleotide does not encode a full length zika virus NS4a.

10. The zika virus-like particle of claim 9, wherein the full length zika virus NS4a comprises a sequence at least 90% identical to SEQ ID NO: 10 or 2942.

11. The zika virus-like particle of any one of claims 1 or 5-10, wherein the polynucleotide encodes a NS4b comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948.

12. The zika virus-like particle of any one of claims 1 or 5-11, wherein the polynucleotide encodes a NS4b comprising a N-terminal disordered region.

13. The zika virus-like particle of claim 12, wherein the polynucleotide does not encode a zika virus NS4b comprising a pTMSl, a pTMS2, a pTMS3, a pTMS4, and a pTMS5.

14. The zika virus-like particle of any one of claims 1, or 5-13, wherein the polynucleotide does not encode a full length zika virus NS4b.

15. The zika virus-like particle of claim 14, wherein the full length zika virus NS4b comprises a sequence at least 90% identical to SEQ ID NO: 12 or 2944.

16. The zika virus-like particle of any one of claims 1 or 5-15, wherein the polynucleotide encodes a zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947.

17. The zika virus-like particle of any one of claims 1 or 5-16, wherein the one or more structural proteins comprises zika virus viral membrane protein (prM / M) or a derivative of the zika virus prM / M, zika virus viral envelope protein (E) or a derivative of the zika virus E, or a combination thereof.

18. The zika virus-like particle of any one of claims 1 or 5-17, wherein the polynucleotide encodes a zika virus capsid protein (C) comprising a sequence at least 90% identical to any one of SEQ ID NOS: 2908-2917.

19. The zika virus-like particle of any one of claims 1 or 5-18, wherein the polynucleotide does not encode a full length zika virus C.

20. The zika virus-like particle of claim 19, wherein the full length zika virus C comprises a sequence at least 90% identical to SEQ ID NO: 3 or 2901.

21. The zika virus-like particle of any one of claims 1 or 5-20, comprising the one or more viral non- structural proteins and the one or more viral structural proteins, optionally wherein the one or more structural proteins are assembled into a zika virus-like shell, and further optionally wherein the zika virus-like particle is not replication competent.

22. A method of producing the zika virus-like particle of any one of claims 1-2 or 5-21, the method comprising introducing into a host cell: the polynucleotide of any one of claims 1-2 or 5-21, a second polynucleotide encoding the one or more viral non- structural proteins, and a third polynucleotide encoding the one or more viral structural proteins.

23. The method of claim 22, wherein each of the second polynucleotide and the third polynucleotide independently comprise a 5’ UTR that is not a zika virus 5’ UTR, and a 3’ UTR that is not a zika virus 3’ UTR.

24. The zika virus-like particle of any one of claims 1-2 or 5-21, or the method of any one of claims 3-4 or 22-23, wherein the zika virus 5’ UTR comprises a sequence at least 90% identical to any one of SEQ ID NOS: 1, 324-633, or the 5’ UTR of the zika nucleotide sequences of Table IB; and wherein the zika virus 3’ UTR comprises a sequence at least 90% identical to any one of SEQ ID NOS: 2, 14-323, or the 3’ UTR of the zika nucleotide sequences of Table IB.

25. A composition comprising: (a) a first polynucleotide encoding a zika virus structural protein, (b) a second polynucleotide encoding a zika virus non- structural (NS) protein, and (c) a third polynucleotide encoding a zika virus 5’ UTR and a zika virus 3’ UTR, wherein the first polynucleotide and the second polynucleotide independently do not encode the zika virus 5’ UTR or the zika virus 3’ UTR.

26. The composition of claim 25, wherein the first polynucleotide encoding the zika virus structural protein comprises:(a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus;(b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus;(c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C;(d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M;(e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E, or(f) a combination of two or more selected from (a) to (e).

27. The composition of claim 25 or claim 26, wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919.

28. The composition of any one of claims 25-27, wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

29. The composition of any one of claims 25-28, wherein the second polynucleotide encoding a zika virus non-structural (NS) protein comprises:(a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus;(b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus;(c) a polynucleotide encoding a zika virus non-structural protein 1 (NSl) or the derivative of the zika virus NSl;(d) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A;(e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A;(f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B;(g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3;(h) a polynucleotide encoding a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A;(i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K;(j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B;(k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS 5; or(l) a combination of two or more selected from (a) to (k).

30. The composition of claim 29, wherein the second polypeptide further comprises a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E.

31. The composition of any one of claims 29-30, wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919.

32. The composition of any one of claims 29-31, wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

33. The composition of any one of claims 25-32, wherein the third polynucleotide encoding a zika virus 5’ UTR of Table IB and a zika virus 3’ UTR of Table IB comprises:(a) a polynucleotide encoding a zika virus 5’ UTR of the zika nucleotide sequences of Table IB;(b) a polynucleotide encoding a zika virus 3’ UTR of the zika nucleotide sequences of Table IB;(c) a polynucleotide of interest;(d) a non- structural protein, or a derivative of a non- structural protein;(e) a structural protein, or a derivative of a structural protein; or(f) a combination of two or more selected from (a) to (e).

34. The composition of claim 33, wherein the non-structural protein comprises a zika virus non- structural protein 4 A (NS4A) or the derivative of the zika virus NS4A, a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, and a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B.

35. The composition of claim 33 or claim 34, wherein the non-structural protein comprises a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A comprising a sequence at least 90% identical to SEQ ID NO: 2929 or 2946, a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947, and a zika virus non- structural protein NS4B (NS4B) or the derivative of the zika virus NS4B comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948.

36. The composition of any one of claims 33-35, wherein the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C.

37. The composition of any one of claims 33-36, wherein the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C comprising a sequence at least 90% identical to SEQ ID NOS: 2908-2917.

38. A method of producing a zika virus like particle comprising introducing into a host cell the composition of any one of claims 25-37.

39. A structural protein plasmid comprising:(a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus;(b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus;(c) a polynucleotide encoding a zika virus capsid protein (C), or a derivative of a zika virus C;(d) a polynucleotide encoding a zika virus viral membrane protein (prM / M) or a derivative of a zika virus prM / M; and(e) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E; wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919, and wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

40. A non-structural protein plasmid comprising:(a) a polynucleotide encoding a 5’ UTR that is not a 5’ UTR of a zika virus or a sequence at least 90% identical to the 5’ UTR of the zika virus;(b) a polynucleotide encoding a 3’ UTR that is not a 3’ UTR of a zika virus or a sequence at least 90% identical to the 3’ UTR of the zika virus;(c) a polynucleotide encoding a zika virus non-structural protein 1 (NSl) or the derivative of the zika virus NSl;(d) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A;(e) a polynucleotide encoding a zika virus non-structural protein 2 A (NS2A) or the derivative of the zika virus NS2A;(f) a polynucleotide encoding a zika virus non-structural protein 2B (NS2B) or the derivative of the zika virus NS2B;(g) a polynucleotide encoding a zika virus non-structural protein 3 (NS3) or the derivative of the zika virus NS3;(h) a polynucleotide encoding a zika virus non-structural protein 4 A (NS4A) or the derivative of the zika virus NS4A;(i) a polynucleotide encoding a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K;(j) a polynucleotide encoding a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B;(k) a polynucleotide encoding a zika virus non-structural protein NS5 (NS5) or the derivative of the zika virus NS 5;(l) a polynucleotide encoding a zika virus viral envelope protein (E) or a derivative of a zika virus E; or(m) a combination of two or more selected from (a) to (1); wherein the 5’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2918 or 2919, and wherein the 3’ UTR of the first polynucleotide comprises a sequence at least 80%, 90%, or 100% identical to SEQ ID NO: 2920 or 2921.

41. A polynucleotide of interest plasmid comprising:(a) a polynucleotide encoding a zika virus 5’ UTR of the zika nucleotide sequences of Table IB;(b) a polynucleotide encoding a zika virus 3’ UTR of the zika nucleotide sequences of Table IB;(c) a polynucleotide of interest;(d) a non-structural protein, or a derivative of a non-structural protein;(e) a structural protein, or a derivative of a structural protein; or(f) a combination of two or more selected from (a) to (e); wherein the non-structural protein comprises a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A, a zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K, and a zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B, and wherein the structural protein comprises a zika virus capsid protein (C), or a derivative of a zika virus C.

42. The polynucleotide of interest plasmid of claim 41, wherein the non-structural protein comprises a zika virus non-structural protein 4A (NS4A) or the derivative of the zika virus NS4A comprising a sequence at least 90% identical to SEQ ID NO: 2929 or 2946, the zika virus non-structural protein 2K (2K) or the derivative of the zika virus 2K comprising a sequence at least 90% identical to SEQ ID NO: 2930 or 2947, and the zika virus non-structural protein NS4B (NS4B) or the derivative of the zika virus NS4B comprising a sequence at least 90% identical to SEQ ID NO: 2931 or 2948.

43. The polynucleotide of interest plasmid of claim 41 or claim 42, wherein the structural protein comprises a zika virus capsid protein (C), or the derivative of a zika virus C comprising a sequence at least 90% identical to SEQ ID NOS: 2908-2917.

44. A method for producing a zika virus-like particle, comprising introducing into a host cell: (a) the structural protein plasmid of claim 39, (b) the non- structural protein plasmid of claim 40, and (c) the polynucleotide of interest plasmid of any one of claims 41-43.

45. A method of treating a disease or condition, comprising administering to a subject in need thereof the zika virus-like particle of any one of claims 1-2, 5-21, 24 or the zika virus-like particle produced by the method of any one of claim 3-4, 22-23, 38, or 44.

46. A method of triggering or boosting an immune response, the method comprising administering to a subject in need thereof an effective amount of the zika virus-like particle of any one of claims 1-2, 5-21, 24, or the zika virus-like particle produced by the method of any one of claim 3-4, 22-23, 38, or 44.

47. A vaccine composition comprising the zika virus-like particle of any one of claims 1-2, 5- 21, 24, or the zika virus-like particle produced by the method of claim 3-4, 22-23, 38, or 44.

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