Nucleic Acid Compositions and Methods of Use for Delivery of Exogenous Polynucleotides - Patent application

Abstract

Disclosed herein are nucleic acid compositions and methods of use for delivery of exogenous polynucleotides.

Description

[Technical field]

[0001] This application claims the benefit of U.S. Patent Application No. 63 / 312,785, filed February 22, 2022, the entirety of which is incorporated herein by reference.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted in XML format, the entirety of which is incorporated herein by reference. The XML copy created on Feb. 21, 2023 is named 61771-703_601_SL.xml and is 2,964,424 bytes in size. [Background technology]

[0003] Cellular delivery of various biologically active compounds, such as antigens and proteins of interest, is of great value in both basic research and industrial applications. However, this is often compromised by two limitations: high toxicity to normal tissues and low delivery efficiency, given that living cells are highly restricted by their complex membrane systems. Particular challenges are seen in delivery efforts targeted to the central nervous system (CNS). Summary of the Invention

[0004] In one aspect, to address the need for new delivery methods to target cells, recombinant Zika viruses and Zika virus-like particles carrying a polynucleotide of interest are provided herein. In some embodiments, the viruses and virus-like particles contain Zika structural proteins. Methods of use include applying such recombinant Zika viruses or such Zika virus-like particles to target cells. In some cases, such methods of use treat disorders of the central nervous system, taking into account its preferred CNS tropism. In some cases, the viruses and virus-like particles do not contain propagating Zika viruses.

[0005] In another aspect, to address the need for new delivery techniques to target cells, provided herein are transcripts of a polynucleotide of interest (e.g., an exogenous polynucleotide) and a nucleic acid composition carrying a regulatory region of the Zika genome. An example method of use includes encapsulating such a transcript with a lipid-based reagent and applying such a compound to a target cell. In some embodiments, the nucleic acid composition includes a Zika 5'UTR, a polynucleotide of interest, and a Zika 3'UTR. In some embodiments, the nucleic acid composition does not encode a Zika structural protein. In some embodiments, the nucleic acid composition does not encode a Zika nonstructural protein. In some embodiments, the polynucleotide of interest has a size of up to about 5700, 5600, 5500, 5400, 5300, 5200, 5100, 5000, 4900, 4800, 4700, 4600, or 4500 nucleotides. In some embodiments, a polynucleotide of interest has a size of about 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 nucleotides up to about 5700, 5600, 5500, 5400, 5300, 5200, 5100, 5000, 4900, 4800, 4700, 4600, or 4500 nucleotides.

[0006] In some embodiments, the nucleic acid composition comprises (i) a polynucleotide encoding a Zika virus capsid protein (C) or a derivative of Zika virus C, a polynucleotide encoding a Zika virus membrane protein (prM / M) or a derivative of Zika virus prM / M, a polynucleotide encoding a Zika virus envelope protein (E) or a derivative of Zika virus E, or any combination thereof, and (ii) a polynucleotide exogenous to the Zika virus. In some embodiments, the nucleic acid composition comprises a derivative of Zika virus C, wherein the derivative encodes at least 5, at least 10, at least 15, at least 20, at least 25, or at least 30, or about 5, about 10, about 15, about 20, about 25, or about 30 amino acids of C. In some embodiments, the nucleic acid composition comprises a derivative of Zika virus C, wherein the derivative encodes about 5, about 10, about 15, about 20, or about 25 to about 30 amino acids of C. In some embodiments, the nucleic acid composition comprises a polynucleotide encoding C. In some embodiments, the nucleic acid composition does not encode a polynucleotide encoding a Zika virus membrane protein (prM / M) or a derivative of Zika virus prM / M, and does not encode a polynucleotide encoding a Zika virus envelope protein (E) or a derivative of Zika virus E. In some embodiments, the exogenous polynucleotide has a size of up to about 5700, 5600, 5500, 5400, 5300, 5200, 5100, 5000, 4900, 4800, 4700, 4600, or 4500 nucleotides. In some embodiments, the exogenous polynucleotide has a size of about 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 nucleotides up to about 5700, 5600, 5500, 5400, 5300, 5200, 5100, 5000, 4900, 4800, 4700, 4600, or 4500 nucleotides. In some embodiments, the nucleic acid composition is a delivery vector for delivering the exogenous polynucleotide to a target cell of a subject. The target cell may be a cell of the central nervous system.

[0007] In some embodiments, the nucleic acid composition comprises a polynucleotide encoding Zika virus C or a derivative of Zika virus C, a polynucleotide encoding Zika virus prM / M or a derivative of Zika virus prM / M, and a polynucleotide encoding Zika virus E or a derivative of Zika virus E. In some embodiments, the polynucleotide encoding Zika virus C or a derivative of Zika virus C, a polynucleotide encoding Zika virus prM / M or a derivative of Zika virus prM / M, and a polynucleotide encoding Zika virus E or a derivative of Zika virus E are expressed on one or more separate nucleic acids.

[0008] In some embodiments, the polynucleotide encoding a derivative of Zika virus C comprises at least one substitution, at least one deletion, and / or at least one insertion compared to wild-type Zika virus C. In some embodiments, the Zika virus C or derivative of Zika virus C comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table ID, Table 1I, Table 2, or Table 3. In some embodiments, the derivative of Zika virus C comprises about 5, about 10, about 15, about 20, or about 25 to about 30 amino acids of a sequence in Table 1A, Table ID, Table 1I, Table 2, or Table 3.

[0009] In some embodiments, a polynucleotide encoding a derivative of Zika virus prM / M comprises at least one substitution, at least one deletion, and / or at least one insertion compared to a wild-type Zika virus prM / M. In some embodiments, the Zika virus prM / M or a derivative of Zika virus prM / M comprises a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1F, or Table 1K.

[0010] In some embodiments, the polynucleotide encoding a derivative of Zika virus E comprises at least one substitution, at least one deletion, and / or at least one insertion compared to wild-type Zika virus E. In some embodiments, the polynucleotide encoding E is translated into wild-type Zika virus E. In some embodiments, the Zika virus E or derivative of Zika virus E comprises a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1E, or Table 1J.

[0011] In some embodiments, the nucleic acid composition comprises a 5' untranslated region (5'UTR) of a Zika virus. In some embodiments, the Zika virus 5'UTR or a derivative of the Zika virus 5'UTR comprises a sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1C, Table 1H, or Table 2.

[0012] In some embodiments, the nucleic acid composition comprises a 3' untranslated region (3'UTR) of a Zika virus. In some embodiments, the Zika virus 3'UTR or a derivative of the Zika virus 3'UTR comprises a sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table IB, Table 1G, or Table 2.

[0013] In some embodiments, the nucleic acid composition does not include a polynucleotide encoding one or more nonstructural (NS) proteins selected from (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) two or more of (i)-(vii). In other embodiments, the nucleic acid composition includes a polynucleotide encoding one or more nonstructural (NS) proteins selected from (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) two or more of (i)-(vii). In some embodiments, NS1 is Zika virus NS1, NS2A is Zika virus NS2A, NS2B is Zika virus NS2B, NS3 is Zika virus NS3, NS4A is Zika virus NS4A, NS4B is Zika virus NS4B, or NS5 is Zika virus NS5, or any combination of two or more of these.

[0014] In some embodiments, the components of the nucleic acid composition include African Zika virus components, Asian Zika virus components, or Brazilian Zika virus components, or combinations thereof. In one particular embodiment, the Zika virus is the African MR766 strain.

[0015] In some embodiments, the exogenous polynucleotide is an antigen or an antigenic epitope thereof. In some embodiments, the antigen or an antigenic epitope thereof is disease associated. In certain embodiments, the antigen or an antigenic epitope thereof is derived from a pathogen.

[0016] In some embodiments, the pathogen is a virus. In certain embodiments, the virus is human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papilloma virus (HPV-6, HPV-11), measles virus, rabies virus, polio virus, or yellow fever virus.

[0017] In some embodiments, the pathogen is a bacteria. In certain embodiments, the bacteria is selected from the group consisting of Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, and the like. 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans.

[0018] In some embodiments, the pathogen is a fungus, an amoeba, or a parasite. In certain embodiments, the fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 These are Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0019] In some embodiments, the exogenous polynucleotide encodes a gene editing tool. In certain embodiments, the gene editing tool is selected from the group consisting of meganuclease-related agents, CRISPR-related agents, TALEN-related agents, and zinc finger-related agents.

[0020] In some embodiments, the exogenous polynucleotide is a small interfering RNA (siRNA), an antisense RNA, a microRNA (miRNA), a small or short hairpin RNA (shRNA), a guide RNA (gRNA), a clustered regularly interspaced short palindromic repeat RNA (crRNA), a trans-activating clustered regularly interspaced short palindromic repeat RNA (tracrRNA), an immunostimulatory oligonucleotide, an antisense nucleic acid, or a ribozyme. In certain embodiments, the exogenous polynucleotide targets beta-secretase 1 (BACE1) and / or amyloid precursor protein (APP).

[0021] In some embodiments, the exogenous polynucleotide encodes a polypeptide associated with a genetic disorder. In certain embodiments, the polypeptide is brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neprilysin inhibitor (NEP), endothelin-converting enzyme (ECE), cathepsin B (CTSB), apolipoprotein E2 (APOE2), SH3 and multiple ankyrin repeat domains protein (SHANK), neurturin (NRTN), glial cell line-derived neurotrophic factor (GDNF), brain dopamine neurotrophic factor (CDNF), vascular endothelial growth factor A (VEGF-A), or aromatic L-amino acid decarboxylase (AADC).

[0022] In some embodiments, the exogenous polynucleotide encodes a therapeutic or diagnostic agent. In certain embodiments, the therapeutic agent is an antibody-based therapeutic agent, a hormone, a cytokine, an inhibitor or antagonist of immune checkpoint regulators, an immune stimulatory molecule, or an agonist of immune costimulatory molecules. In further embodiments, the inhibitor or antagonist of immune checkpoint regulators is an anti-PD1 antibody. In another embodiment, 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 yet other embodiments, the cytokine is a lymphokine, a monokine, a polypeptide hormone, a growth hormone, a parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, a glycoprotein hormone, a follicle stimulating hormone (FSH), a thyroid stimulating hormone (TSH), a luteinizing hormone (LH), a hepatocyte growth factor, a fibroblast growth factor, a prolactin, a placental lactogen, a tumor necrosis factor-α and -β, a Mullerian inhibitory substance, a mouse gonadotropin-related peptide, an inhibin, an activin, a vascular endothelial growth factor, an integrin, a thrombopoietin (TPO), These include nerve growth factor, NGF-β, platelet growth factor, transforming growth factor (TGF), TGF-α, TGF-β, insulin-like growth factor-I and -II, erythropoietin (EPO), osteoinductive factor, interferon, e.g., interferon-α, -β, and -γ, colony stimulating factor (CSF), macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF), granulocyte CSF (GCSF), interleukin (IL), tumor necrosis factor, TNF-α, TNF-β, LIF, or Kit ligand (KL), or a combination of two or more thereof.

[0023] In some embodiments, the exogenous polynucleotide is about, or up to, about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length. For example, the exogenous polynucleotide is about 50 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length.

[0024] In some embodiments, the components of the nucleic acid composition described herein are expressed on one nucleic acid or two or more separate nucleic acids.In some embodiments, the nucleic acid composition further comprises one or more expression control elements in operable linkage that cause the expression of the nucleic acid composition in vitro or in vivo.In some embodiments, the expression control element is a promoter that drives the expression of the nucleic acid complex in vitro.In certain embodiments, the promoter is T7, T3, SP6, or any phage promoter.

[0025] In some embodiments, the expression control element is a promoter that drives expression of the nucleic acid complex in the target cell. In some specific embodiments, the promoter is CMV, SV40, or any eukaryotic promoter. In some embodiments, the target cell is a neuronal cell, or a non-neuronal cell, VERO, COS, CHO, C6 / 36, HeLa, HEK, HepG2. In some embodiments, the target cell is an oligodendrocyte, a microglia, or an astrocyte.

[0026] In another aspect, provided herein is a pharmaceutical composition comprising a nucleic acid composition described herein, or a pharma- ceutically acceptable salt or derivative thereof.

[0027] In another aspect, provided herein is a recombinant Zika virus or Zika virus-like particle generated from expressing a nucleic acid composition described herein in a producer cell further infected with a second Zika virus, such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus.

[0028] In some embodiments, the second Zika virus is a wild type Zika virus. In some embodiments, the wild type Zika virus is an African strain, an Asian strain, and a Brazilian strain. In other embodiments, the second Zika virus is a modified Zika virus. In some specific embodiments, the modified Zika virus comprises one or more microRNA-based gene silencing mechanisms. In some specific embodiments, the one or more microRNA-based gene silencing mechanisms control the replication of the virus.

[0029] In another aspect, provided herein is a recombinant Zika virus or Zika virus-like particle generated from expressing a nucleic acid composition described herein in a producer cell that is not infected with a second Zika virus, wherein Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0030] In some embodiments, producer cells are Vero E6, HEK, HEK 293T, HEK 293TT, FreeStyle™ 293-F cells, HEK-293.2sus, or C6 / 36 cells.

[0031] In some embodiments, the recombinant Zika virus is replication competent. In other embodiments, the recombinant Zika virus is unable to replicate without reducing vector titer or compromising expression of the exogenous polynucleotide. In some embodiments, the recombinant Zika virus reduces insertional mutagenesis. In other embodiments, the recombinant Zika virus reduces the immune response.

[0032] In another aspect, provided herein is a pharmaceutical composition comprising a recombinant Zika virus or Zika virus-like particle described herein, or a pharma- ceutical acceptable salt or derivative thereof.

[0033] In another aspect, provided herein is a method of delivering an exogenous polynucleotide to a target cell, the method comprising administering to the target cell a recombinant Zika virus or Zika virus-like particle described herein comprising an exogenous polynucleotide.

[0034] In some embodiments, the target cell is a neuronal cell or a non-neuronal cell. In certain embodiments, the neuronal cell is an oligodendrocyte, a microglia, or an astrocyte. In another specific embodiment, the non-neuronal cell is a prostate epithelial cell, a urethral epithelial cell, a Sertoli cell, a Leydig cell, a spermatogonium cell, or a retinal cell.

[0035] In some embodiments, the methods are performed in vitro, ex vivo, or in vivo.

[0036] In some embodiments, the target cells express the exogenous polynucleotide transiently after delivery, hi other embodiments, the target cells express the exogenous polynucleotide persistently after delivery.

[0037] In another aspect, provided herein is a method of treating Alzheimer's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding brain-derived neurotrophic factor (BDNF).

[0038] In another aspect, provided herein is a method of treating autism in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein, the recombinant Zika virus or Zika virus-like particle comprising a polynucleotide encoding an SH3 and multiple ankyrin repeat domains protein (SHANK).

[0039] In yet another aspect, provided herein is a method of treating Parkinson's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding aromatic L-amino acid decarboxylase (AADC).

[0040] In some embodiments, the administering is by systemic delivery.

[0041] In some embodiments, the administering step is performed intravenously and / or intratumorally.

[0042] In some embodiments, the administering step targets ventricular, cisternal, subpial, and / or intrathecal cerebrospinal fluid, hi other embodiments, the administering step is not performed intraparenchymal.

[0043] In some embodiments, the nucleic acid composition comprises (i) a polynucleotide encoding a ribozyme, (ii) a 5' untranslated region (UTR) of Zika virus, (iii) a 3' UTR of Zika virus, and (iv) a polynucleotide exogenous to Zika virus. In some embodiments, the ribozyme is a hammerhead ribozyme or a Hepatitis D 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 D virus (HDV) ribozyme. In some embodiments, the polynucleotide encoding the ribozyme encodes a hammerhead ribozyme and the polynucleotide encoding the second ribozyme encodes a HDV ribozyme. In some embodiments, the nucleic acid composition further comprises a C moiety. A C portion can refer to a polynucleotide that encodes a portion of C, e.g., about 5, about 10, about 15, about 20, or about 25 amino acids to about 30 amino acids of C. In some embodiments, the exogenous polynucleotide has a size of up to about 5700, 5600, 5500, 5400, 5300, 5200, 5100, 5000, 4900, 4800, 4700, 4600, or 4500 nucleotides. In some embodiments, the exogenous polynucleotide has a size of about 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 nucleotides up to about 5700, 5600, 5500, 5400, 5300, 5200, 5100, 5000, 4900, 4800, 4700, 4600, or 4500 nucleotides. In some embodiments, the nucleic acid composition is a delivery vector for delivering the exogenous polynucleotide to a target cell of a subject. The target cell may be a cell of the central nervous system.

[0044] In some embodiments, the Zika virus is an African Zika virus, an Asian Zika virus, a component of a Brazilian Zika virus, or a combination of one or two of these. In certain embodiments, the Zika virus is the African MR766 strain.

[0045] In some embodiments, the exogenous polynucleotide is an antigen or an antigenic epitope thereof. In some embodiments, the antigen or an antigenic epitope thereof is disease associated. In certain embodiments, the antigen or an antigenic epitope thereof is derived from a pathogen.

[0046] In some embodiments, the pathogen is a virus. In certain embodiments, the virus is human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papilloma virus (HPV-6, HPV-11), measles virus, rabies virus, polio virus, or yellow fever virus.

[0047] In some embodiments, the pathogen is a bacteria. In certain embodiments, the bacteria is selected from the group consisting of Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, and the like. 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans.

[0048] In some embodiments, the pathogen is a fungus, an amoeba, or a parasite. In certain embodiments, the fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 These are Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0049] In some embodiments, the exogenous polynucleotide encodes a gene editing tool. In certain embodiments, the gene editing tool is selected from the group consisting of meganuclease-related agents, CRISPR-related agents, TALEN-related agents, and zinc finger-related agents.

[0050] In some embodiments, the exogenous polynucleotide is a small interfering RNA (siRNA), an antisense RNA, a microRNA (miRNA), a small or short hairpin RNA (shRNA), a guide RNA (gRNA), a clustered regularly interspaced short palindromic repeat RNA (crRNA), a trans-activating clustered regularly interspaced short palindromic repeat RNA (tracrRNA), an immunostimulatory oligonucleotide, an antisense nucleic acid, or a ribozyme. In certain embodiments, the exogenous polynucleotide targets beta-secretase 1 (BACE1) and / or amyloid precursor protein (APP).

[0051] In some embodiments, the exogenous polynucleotide encodes a polypeptide associated with a genetic disorder. In certain embodiments, the polypeptide is brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neprilysin inhibitor (NEP), endothelin-converting enzyme (ECE), cathepsin B (CTSB), apolipoprotein E2 (APOE2), SH3 and multiple ankyrin repeat domains protein (SHANK), neurturin (NRTN), glial cell line-derived neurotrophic factor (GDNF), brain dopamine neurotrophic factor (CDNF), vascular endothelial growth factor A (VEGF-A), or aromatic L-amino acid decarboxylase (AADC).

[0052] In some embodiments, the exogenous polynucleotide encodes a therapeutic or diagnostic agent. In certain embodiments, the therapeutic agent is an antibody-based therapeutic agent, a hormone, a cytokine, an inhibitor or antagonist of immune checkpoint regulators, an immune stimulatory molecule, or an agonist of immune costimulatory molecules. In further embodiments, the inhibitor or antagonist of immune checkpoint regulators is an anti-PD1 antibody. In other 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 yet other embodiments, the cytokine is a lymphokine, a monokine, a polypeptide hormone, a growth hormone, a parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, a glycoprotein hormone, a follicle stimulating hormone (FSH), a thyroid stimulating hormone (TSH), a luteinizing hormone (LH), a hepatocyte growth factor, a fibroblast growth factor, a prolactin, a placental lactogen, a tumor necrosis factor-α and -β, a Mullerian inhibitory substance, a mouse gonadotropin-related peptide, an inhibin, an activin, a vascular endothelial growth factor, an integrin, a thrombopoietin (TPO), These include nerve growth factor, NGF-β, platelet growth factor, transforming growth factor (TGF), TGF-α, TGF-β, insulin-like growth factor-I and -II, erythropoietin (EPO), osteoinductive factor, interferon, e.g., interferon-α, -β, and -γ, colony stimulating factor (CSF), macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF), granulocyte CSF (GCSF), interleukin (IL), tumor necrosis factor, TNF-α, TNF-β, LIF, or Kit ligand (KL), or a combination of two or more thereof.

[0053] In some embodiments, the exogenous polynucleotide is about, or up to, about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length. For example, the exogenous polynucleotide is about 50 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length.

[0054] In some embodiments, the nucleic acid composition further comprises one or more expression control elements in operable linkage that cause 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 certain embodiments, the promoter is T7, T3, SP6, or any phage promoter.

[0055] In another aspect, provided herein is a pharmaceutical composition comprising a nucleic acid composition described herein, or a pharma- ceutically acceptable salt or derivative thereof.

[0056] In another aspect, provided herein is a method of delivering an exogenous polynucleotide to a target cell, the method comprising applying to the target cell a compound generated from a nucleic acid composition described herein that includes the exogenous polynucleotide.

[0057] In some embodiments, the compound is produced by encapsulating the transcript produced from the nucleic acid composition described herein with a lipid-based agent. In certain embodiments, the transcript is uncapped. In another particular embodiment, the transcript is produced by transcribing the nucleic acid composition in vitro.

[0058] In some embodiments, the lipid-based agent is a lipofectamine-related reagent, a liposome, or a lipid nanoparticle.

[0059] In another aspect, provided herein is a method of inducing or boosting an immune response in a subject, the method comprising administering to the subject an effective amount of a compound generated from a nucleic acid composition described herein that comprises an antigen or an epitope of the antigen.

[0060] In some embodiments, the compound is produced by encapsulating the transcript produced from the nucleic acid composition described herein with a lipid-based agent. In certain embodiments, the transcript is uncapped. In another particular embodiment, the transcript is produced by transcribing the nucleic acid composition in vitro.

[0061] In some embodiments, the lipid-based agent is a lipofectamine-related reagent, a liposome, or a lipid nanoparticle.

[0062] In some embodiments, the administering step is performed intramuscularly.

[0063] In some embodiments, the antigen or antigenic epitope thereof is derived from a pathogen.

[0064] In some embodiments, the pathogen is a virus. In certain embodiments, the virus is human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papilloma virus (HPV-6, HPV-11), measles virus, rabies virus, polio virus, or yellow fever virus.

[0065] In some embodiments, the pathogen is a bacteria. In certain embodiments, the bacteria is selected from the group consisting of Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, and the like. 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans.

[0066] In some embodiments, the pathogen is a fungus, an amoeba, or a parasite. In certain embodiments, the fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 These are Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0067] Certain embodiments include a method of delivering a polynucleotide to a target cell of a subject, the method comprising administering to the subject a Zika virus or Zika virus-like particle comprising a polynucleotide, where the polynucleotide is not a Zika virus polynucleotide, and the target cell is a cell of the central nervous system. Certain embodiments include a method of delivering a polynucleotide to a subject, the method comprising administering to the subject a Zika virus or Zika virus-like particle comprising a polynucleotide, where the polynucleotide is not a Zika virus polynucleotide, and the Zika virus or Zika virus-like particle does not comprise a proliferating Zika virus. Certain embodiments include a method of delivering a polynucleotide to a subject, the method comprising administering to the subject a Zika virus or Zika virus-like particle comprising a polynucleotide, where the polynucleotide is not a Zika virus polynucleotide, and the polynucleotide has a length of up to about 10,000 nucleotides. In some embodiments, the subject has a disease or disorder of the central nervous system. In some embodiments, the polynucleotide has a length of more than about 6,000 nucleotides. In some embodiments, the subject is a human or a non-human animal.

[0068] Certain embodiments include a Zika virus or Zika virus-like particle comprising a polynucleotide exogenous to the Zika virus having a length of about 6,000 nucleotides to about 10,000 nucleotides, optionally wherein the polynucleotide does not encode a full-length viral structural protein or a full-length viral nonstructural protein. In some embodiments, the Zika virus or Zika virus-like particle of claim 7 is generated from a nucleic acid comprising the polynucleotide. In some embodiments, the nucleic acid further comprises a Zika virus 5'UTR and a Zika virus 3'UTR. In some embodiments, the Zika virus 5'UTR is at least 90% homologous or identical to a Zika virus 5'UTR of Table 2, Table 1A, Table 1C, or Table 1H. In some embodiments, the Zika virus 3'UTR is at least 90% homologous or identical to a Zika virus 3'UTR of Table 2, Table 1A, Table 1B, or Table 1G. In some embodiments, the nucleic acid further comprises a polynucleotide encoding a first ribozyme. In some embodiments, the nucleic acid further comprises a polynucleotide encoding a second ribozyme. In some embodiments, the nucleic acid comprises a polynucleotide encoding a derivative of Zika virus C, the derivative of Zika virus C being a truncation of about 5, about 10, about 15, about 20, about 25, or about 30, or less than about 5, about 10, about 15, about 20, about 25, or about 30 amino acids of Zika virus C. In some embodiments, the nucleic acid does not comprise a polynucleotide encoding a Zika virus capsid protein (C) or a derivative of Zika virus C. In some embodiments, the nucleic acid does not comprise a polynucleotide encoding a Zika virus membrane protein (prM / M) or a derivative of Zika virus prM / M. In some embodiments, the nucleic acid does not comprise a polynucleotide encoding a Zika virus envelope protein (E) or a derivative of Zika virus E.In some embodiments, the nucleic acid does not comprise a polynucleotide encoding one or more nonstructural (NS) proteins, i.e., (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) two or more of (i)-(vii). In some embodiments, the nucleic acid further comprises a promoter. In some embodiments, the nucleic acid comprises a sequence of Table 1A-1K, Table 2, or Table 3.

[0069] Certain embodiments include nucleic acid compositions comprising: (i) a 5' untranslated region (5'UTR) of a first Zika virus; (ii) a 3'UTR of a second Zika virus, the first Zika virus optionally being identical to the second Zika virus; and (iii) a polynucleotide exogenous to the first Zika virus and the second Zika virus. In some embodiments, the polynucleotide is about 5000 bases to about 10000 bases in length. In some embodiments, the nucleic acid does not comprise a polynucleotide encoding a Zika virus envelope protein (E) or a derivative of Zika virus E, and the nucleic acid does not comprise a polynucleotide encoding one or more nonstructural (NS) proteins, i.e., (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) two or more of (i)-(vii). In some embodiments, the nucleic acid composition comprises a sequence of Tables 1A-1K, Table 2, or Table 3. In some embodiments, the polynucleotides exogenous to the first Zika virus and the second Zika virus are exogenous polynucleotides described herein. [Brief description of the drawings]

[0070] 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 (herein referred to as "Figure and FIG.").

[0071] [Figure 1] FIG. 1A-E show diagrams of exemplary replicons described in this disclosure with different example applications. FIG. 1A shows a replicon structure containing a vector or construct, or mRNA, as a gene expressed without capping. The replicon structure may contain C portion proteins. The replicon contains two report genes (GFP and Nluc) for proof-of-concept assays. FIG. 1B shows four possibilities for encapsulating the replicon during delivery. Option 1 is encapsulation with liposomes or nanoparticles for use as a vaccine platform. To maintain natural tropism of the CNS, the replicon is derived from a eukaryotic expression vector transfection (option 4) containing a wild-type (option 2), or modified Zika virus (option 3), or a second construct containing other structural ZIKV proteins. FIG. 1C shows encapsulation options 2 and 3, containing both the replicon and Zika virus genome, including recombinant Zika virus that can be used as an anti-cancer therapy. Figure 1D shows encapsulation options 2 and 3 that contain only the replicon (no Zika virus genome) including virus-like particles that can be used as viral vectors for CNS gene therapy. Figure 1D shows virus-like particles that contain only the replicon RNA resulting from eukaryotic expression vector transfection (option 4). This encapsulation process is non-viral. [Diagram 2] FIG. 2 shows a diagram of another exemplary replicon described in this disclosure compared to the wild-type Zika virus genome. [Diagram 3] FIG. 3 shows exemplary imaging results of cells transfected with a plasmid carrying a GFP construct. [Figure 4] Figure 4A-F show a workflow flow chart for generating recombinant Zika virus by infecting construct-expressing cells with wild-type Zika virus and bright-field microscopy results. [Diagram 5]FIG. 5 shows the plasmid constructs containing Zika virus hammerhead ribozyme, (ii) Zika virus 5' untranslated region (UTR), (iii) Zika virus C portion, and (iv) Zika virus 3'UTR, (v) Zika virus hepatitis D virus (HDV) ribozyme, and two reporter genes (eGFP and Nluc). [Figure 6] FIG. 6 shows a flow chart of the RNA transfection of in vitro transcripts and the quantification of the expression of the exogenous reporter gene Nluc in the cell lysates or supernatants of transfected cells. [Figure 7] Figure 7 shows the generation of exogenous genes (steps 1-3) and their delivery and expression in Daoy and Vero cells using Zika virus (step 4). [Figure 8] Figure 8 shows RT PCR from samples 5 days after infection of VERO cells with the ZIKV viral vector. Quantitation of RNA copy numbers present in the viral vector (pREP+ZIKV) and ZIKV (subjected to ZIKV infection only) is compared using pREP-specific TaqMan primers (black bars) and ZIKV (gray bars). [Figure 9] Figure 9 shows the tropism of ZIKVV in different types of cells and the expression of the NanoNuc reporter gene delivered by pREP. The control group had no virus infection, the ZIKV wild-type group was infected with wild-type ZIKA virus, and the ZIKVV group was infected with the novel ZIKA virus vector. [Figure 10] FIG. 10 is an example of a construct containing polycistronic luciferase and β-galactosidase genes within the Zika virus 5′UTR and Zika virus 3′UTR. [Figure 11] FIG. 11 is an example of a construct containing genes encoding SARS-CoV-2 proteins within the Zika virus 5'UTR and Zika virus 3'UTR. [Figure 12]Figure 12 presents a graph showing quantification of luciferase and β-galactosidase in Vero cells after transfection with the pREP-Luc / Gal vector compared to control cells (transfected with Lipofectamine only). High levels of expression of NanoLuc and β-galactosidase confirm the ability of the ZIKV construct to deliver multiple genes. [Figure 13] FIG. 13 shows ELISA results for detection of recombinant SARS-CoV-2 spike protein produced in Vero cells after transfection with a pREP mRNA construct containing a gene of interest that encodes the spike protein. [Figure 14A] Figures 14A-14E are examples of constructs with partial capsid genes of different lengths: 15 bp (Figure 14A), 30 bp (Figure 14B), 45 bp (Figure 14C), 60 bp (Figure 14D), and 75 bp (Figure 14E). [Figure 14B] Figures 14A-14E are examples of constructs with partial capsid genes of different lengths: 15 bp (Figure 14A), 30 bp (Figure 14B), 45 bp (Figure 14C), 60 bp (Figure 14D), and 75 bp (Figure 14E). [Figure 14C] Figures 14A-14E are examples of constructs with partial capsid genes of different lengths: 15 bp (Figure 14A), 30 bp (Figure 14B), 45 bp (Figure 14C), 60 bp (Figure 14D), and 75 bp (Figure 14E). [Figure 14D] Figures 14A-14E are examples of constructs with partial capsid genes of different lengths: 15 bp (Figure 14A), 30 bp (Figure 14B), 45 bp (Figure 14C), 60 bp (Figure 14D), and 75 bp (Figure 14E). [Figure 14E] Figures 14A-14E are examples of constructs with partial capsid genes of different lengths: 15 bp (Figure 14A), 30 bp (Figure 14B), 45 bp (Figure 14C), 60 bp (Figure 14D), and 75 bp (Figure 14E). [Figure 15A]Figures 15A-15B show the biodistribution and tropism of ZIKVV in mice 6 hours after IP injection of ZIKVV (Figure 15A) and 48 hours after IP injection of ZIKVV (Figure 15B). [Figure 15B] Figures 15A-15B show the biodistribution and tropism of ZIKVV in mice 6 hours after IP injection of ZIKVV (Figure 15A) and 48 hours after IP injection of ZIKVV (Figure 15B). [Figure 16A] Figure 16A shows the encapsulation efficiency of pREP RNA in all the tested plasmids (Figures 14A-14E have C moieties of 15 bp, 30 bp, 45 bp, and 60 bp, respectively). The encapsulation efficiency of pREP RNA is higher in the plasmids with C moieties of 15 bp, 30 bp, 45 bp, and 60 bp, indicating that ZIKVV carrying pREP RNA with a capsid having 20 or fewer amino acids is efficiently encapsulated by the viral vector. [Figure 16B] FIG. 16B shows that encapsulation resulting from DNA transfection was efficient, and the plasmid with a 45 bp C portion had a higher amount of pREP RNA. [Figure 17A] Figures 17A-17B show the quantification of Nanoluc in Vero cells after contact with viral vectors carrying 5, 10, 15, 20, and 25 amino acids (pREP+ZIKV) from both RNA (Figure 17A) and DNA (Figure 17B) transfections compared to ZIKV (ZIKV infection only). Higher levels of NanoLuc expression are demonstrated in viral vectors of all capsid sequence sizes compared to the control group. These results show that ZIKVV can encapsulate pREP RNA at the C portion (15, 30, 45, 60, and 75 bp) and confirm that NanoLuc is delivered by reporter gene expression in target cells. [Figure 17B]Figures 17A-17B show the quantification of Nanoluc in Vero cells after contact with viral vectors carrying 5, 10, 15, 20, and 25 amino acids (pREP+ZIKV) from both RNA (Figure 17A) and DNA (Figure 17B) transfections compared to ZIKV (ZIKV infection only). Higher levels of NanoLuc expression are demonstrated in viral vectors of all capsid sequence sizes compared to the control group. These results show that ZIKVV can encapsulate pREP RNA at the C portion (15, 30, 45, 60, and 75 bp) and confirm that NanoLuc is delivered by reporter gene expression in target cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0072] Various nucleic acid compositions comprising an exogenous polynucleotide are provided herein. The nucleic acid composition may be useful for delivering the exogenous polynucleotide to a target cell. The target cell may be a cell of the central nervous system. The target cell may be present in a subject, and the nucleic acid composition delivers the exogenous polynucleotide to the target cell of the subject. Certain nucleic acid compositions include an exogenous polynucleotide up to about 6,000 or 10,000 nucleotides in length, for example, from about 5 nucleotides to about 6,000 nucleotides in length. The nucleic acid composition may include a 5' untranslated region (UTR) of a Zika virus and a 3' UTR of a Zika virus, optionally not including a polynucleotide encoding one or more nonstructural proteins of the virus, and further optionally not including a polynucleotide encoding one or more structural proteins of the virus.

[0073] In one aspect, provided herein is a nucleic acid composition comprising (i) a polynucleotide encoding a ribozyme, (ii) a 5' untranslated region (UTR) of a Zika virus, (iii) a 3' UTR of a Zika virus, and (iv) a polynucleotide exogenous to a Zika virus. In some embodiments, the ribozyme is a hammerhead ribozyme or a Hepatitis D 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 D virus (HDV) ribozyme. In some embodiments, the polynucleotide encoding the ribozyme encodes a hammerhead ribozyme and the polynucleotide encoding the second ribozyme encodes a HDV ribozyme. In some embodiments, the nucleic acid composition further comprises a C moiety. For example, the C moiety is a derivative of C, where the derivative encodes a truncation of C (e.g., about 30, about 25, about 20, about 15, about 10, or about 5, or less than about 30, about 25, about 20, about 15, about 10, or about 5 amino acids).

[0074] In one aspect, provided herein is a nucleic acid composition comprising a sequence encoding one of the structural proteins from a Zika virus or a derivative thereof having an exogenous polynucleotide. As a non-limiting example, the nucleic acid composition comprises a derivative of the Zika capsid protein (C), wherein the derivative encodes a truncation of C (e.g., about 30, about 25, about 20, about 15, about 10, or about 5 amino acids, or less than about 30, about 25, about 20, about 15, about 10, or about 5 amino acids).

[0075] Non-limiting example nucleic acids herein may include one or more sequences from Tables 1A-1K, Table 2, and Table 3.

[0076] Further provided herein is a recombinant Zika virus or Zika virus-like particle comprising all three structural proteins from Zika virus or a derivative thereof (a polynucleotide encoding Zika virus capsid protein (C) or a derivative of Zika virus C, a polynucleotide encoding Zika virus membrane protein (prM / M) or a derivative of Zika virus prM / M, a polynucleotide encoding Zika virus envelope protein (E) or a derivative of Zika virus E). Further provided herein is a method of using such a recombinant Zika virus or such a Zika virus-like particle to deliver an exogenous polynucleotide to a target cell. In a preferred embodiment, such a recombinant Zika virus effectively crosses the blood-brain barrier and has a tropism to target the central nervous system (CNS). Thus, in a preferred embodiment, such a recombinant Zika virus is used to deliver an exogenous polynucleotide to the CNS.

[0077] Before describing the method and composition of the present invention, it should be understood that the present disclosure is not limited to the specific method or composition described, and therefore can be modified.Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art.All publications mentioned herein are incorporated by reference to disclose and describe in relation to the method and / or material in which the publication is cited.It will be understood that the present disclosure supersedes the disclosure of any of the incorporated publications to the extent of any discrepancy.

[0078] As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has distinct components and features which may be readily separated from or combined with the features of any of the other embodiments without departing from the scope or spirit of the disclosure. Any recited method may be carried out in the order of events recited or in any other order which is logically possible.

[0079] Where a range of values ​​is provided, it is understood that each intervening value between the upper and lower limits of that range, to the first decimal place of the lower limit, is also specifically disclosed unless otherwise expressly stated. Each smaller range between any stated value or intervening value of the stated range is encompassed herein. The upper and lower limits of these smaller ranges may be independently included or excluded within the range, and if either or neither of the limits are included within these smaller ranges, or both are included, each range is encompassed herein, subject to any specifically excluded limit within the stated range. When a stated range includes one or both limits, ranges excluding either or both of those included limits are also included.

[0080] The term "about" or "approximately" means within an acceptable error range of a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, "about" may, by convention, mean within or more than one standard deviation of a given value. When a particular value is described in this application and claims, unless otherwise indicated, 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."

[0081] The terms "individual," "patient," or "subject" may be used interchangeably. None of the terms require or are limited to a situation characterized by the supervision (e.g., continuous or intermittent) of a medical professional (e.g., a physician, registered nurse, nurse practitioner, physician assistant, floor staff, or hospice personnel). In some embodiments, a patient, subject, or individual may be under the supervision of a medical professional.

[0082] As used herein, the term "heterologous nucleic acid sequence" or "exogenous nucleic acid sequence" or "transgene" in reference to a particular virus may refer to a nucleic acid sequence derived from a source other than the particular virus.

[0083] The terms "inhibiting," "reducing," or "prevention," or any variation of these terms, as referred to herein, may include any measurable reduction or complete inhibition to achieve a desired result.

[0084] As used herein, a "promoter" may be a control sequence that is a region of a nucleic acid sequence where the initiation and rate of transcription are controlled. In certain embodiments, a promoter may contain genetic elements to which regulatory proteins and molecules, such as RNA polymerase and other transcription factors, can bind. The terms "operably arranged," "operably linked," "under control," and "under transcriptional control" may mean that the promoter is in the correct functional location and / or orientation relative to a nucleic acid sequence to control the transcription 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.

[0085] The term "percentage identical" as used herein may be a calculation of the "homology" or "percent homology" of two or more nucleotide or amino acid sequences, which may be determined by aligning the sequences for optimal comparison purposes (e.g., gaps may be introduced into the sequence of the 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 (i.e., % homology = number of identical positions / total number of positions x 100). For example, a position in a first sequence may be occupied by the same nucleotide as the corresponding position in a second sequence, in which case the molecules are identical at that position. The percent homology between two sequences may be a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced to optimally align the two sequences and the length of each gap. In some embodiments, the length of the aligned sequences 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. The homology between two sequences may be determined by a BLAST® search. The homology may be between the full length of the two sequences or between a fraction of the full length of the two sequences. The two sequences may be genes, nucleotide sequences, protein sequences, peptide sequences, amino acid sequences, or fragments thereof. The actual comparison of the two sequences may be accomplished by known methods, for example, using a mathematical algorithm. Non-limiting examples of such mathematical algorithms may be found in Karlin, S. and Altschul, S., Proc. Natl. Acad. Sci. USA, 90-5873-5877 (1993). Such algorithms 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 altered (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 achieved, for example, using the GAP program in the GCG software package (Accelrys, Cambridge, UK).

[0086] The term "subject" may refer to an animal, including, but not limited to, a primate (e.g., a human), cow, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. The terms "subject" and "patient" are used interchangeably herein in reference to a mammalian subject, e.g., a human subject or a non-human subject.

[0087] The terms "treat," "treating," and "treatment" may be meant to include alleviating or arresting a disorder, disease, or condition, or one or more symptoms associated with a disorder, disease, or condition, or alleviating or eradicating the cause of the disorder, disease, or condition itself. Desirable effects of treatment may include, but are not limited to, prevention of disease onset or recurrence, alleviation of symptoms, attenuation of any direct or indirect pathological consequence of the disease, prevention of metastasis, reducing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.

[0088] The term "therapeutically effective amount" may refer to an amount of a compound that, when administered, may be sufficient to prevent the progression 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" may also refer to an amount of a compound sufficient to elicit the biological or medical response in a cell, tissue, system, animal, or human that is being sought by a researcher, veterinarian, physician, or clinician.

[0089] The terms "pharmaceutically acceptable carrier", "pharmaceutically acceptable excipient", "physiologically acceptable carrier", or "physiologically acceptable excipient" may refer to a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating material. An ingredient may be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of a pharmaceutical formulation. An ingredient may also be suitable for use in contact with the tissues or organs of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity, or other problem or complication, commensurate with a reasonable benefit-risk ratio. See Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 5th Edition, edited by Rowe et al., The Pharmaceutical Press and the American Pharmaceutical Association: 2005; and Handbook of Pharmaceutical Additives, 3rd Edition, edited by Ash and Ash, Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, edited by Gibson, CRC Press LLC: Boca Raton, FL, 2004.

[0090] The term "pharmaceutical composition" may refer to a mixture of a compound disclosed herein with other chemical components, such as diluents or carriers. A pharmaceutical composition can facilitate administration of a compound to an organism. There are multiple techniques of administering a compound in the art, including but not limited to oral, injection, aerosol, parenteral, and topical administration. A pharmaceutical composition can also be obtained by reacting a compound with an inorganic or organic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0091] The term "Zika virus-like particle" may refer to nanoscale multiprotein structures composed of assembled Zika virus proteins that mimic the organization and conformation of the Zika virus, but do not contain viral genetic material and are therefore non-infectious. Zika virus-like particles may closely resemble the Zika virus.

[0092] In some embodiments, a "derivative" of a polypeptide or polynucleotide, as used herein, refers to a sequence that is at least 80% identical to the polypeptide or polynucleotide, respectively. In some embodiments, a "derivative" of a polypeptide or polynucleotide, as used herein, refers to a sequence that is at least 80% homologous to the polypeptide or polynucleotide, respectively. In some embodiments, a "derivative" of a polypeptide or polynucleotide, as used herein, refers to a sequence that has or has 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.

[0093] Nucleic acid composition In one aspect, provided herein is a nucleic acid composition comprising: (i) a polynucleotide encoding a Zika virus capsid protein (C) or a derivative of Zika virus C, a polynucleotide encoding a Zika virus membrane protein (prM / M) or a derivative of Zika virus prM / M, a polynucleotide encoding a Zika virus envelope protein (E) or a derivative of Zika virus E, or any combination thereof; and (ii) a polynucleotide exogenous to Zika virus.

[0094] In another aspect, provided herein is a nucleic acid composition comprising a Zika virus 5'UTR, a Zika virus 3'UTR, and a polynucleotide exogenous to Zika virus. In some embodiments, the nucleic acid does not comprise a polynucleotide encoding a Zika virus capsid protein (C) or a derivative of Zika virus C, a polynucleotide encoding a Zika virus membrane protein (prM / M) or a derivative of Zika virus prM / M, a polynucleotide encoding a Zika virus envelope protein (E) or a derivative of Zika virus E, or any combination thereof.

[0095] Structural and nonstructural proteins from Zika virus The wild-type Zika virus genome contains 10.8 kilobases of single-stranded positive-stranded RNA encoding three structural proteins (C, prM / M, and envelope E) and seven nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5). In addition, short UTRs are present at both the 5' and 3' ends of the genome. Various embodiments are provided herein for generating recombinant Zika viruses that contain at least one or all of the three structural proteins. In some embodiments, a nucleic acid for generating a recombinant Zika virus is provided, where the nucleic acid lacks a polynucleotide encoding one or more or all of the Zika virus structural proteins and the Zika virus nonstructural proteins. The nucleic acid may include the Zika virus 5'UTR and the Zika virus 3'UTR, and an exogenous nucleic acid.

[0096] In some embodiments, the nucleic acid composition comprises a polynucleotide encoding Zika virus C, a polynucleotide encoding Zika virus prM / M, and a polynucleotide encoding Zika virus E. When three structural proteins are expressed as nucleic acid compositions described herein, in some embodiments, these structural proteins may be expressed on one nucleic acid. When three structural proteins are expressed as nucleic acid compositions described herein, in other embodiments, these structural proteins may be expressed on more than one nucleic acid.

[0097] In some embodiments, the polynucleotide encoding a derivative of Zika virus C comprises at least one substitution, at least one deletion, and / or at least one insertion compared to wild-type Zika virus C. In certain embodiments, the Zika virus C or derivative of Zika virus C comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table ID, Table 1I, Table 2, or Table 3. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus C that encodes an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the wild-type C sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus C that encodes an amino acid sequence that is at least 95% identical to the wild-type C sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus C that encodes an amino acid sequence that is at least 96% identical to the wild-type C sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus C that encodes an amino acid sequence that is at least 97% identical to the wild-type C sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus C that encodes an amino acid sequence that is at least 98% identical to the wild-type C sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus C that encodes an amino acid sequence that is at least 99% identical to the wild-type C sequence. In some embodiments, the nucleic acid composition comprises a derivative of Zika virus C that encodes about 5 to about 30 amino acids of a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1D, Table 1I, Table 2, Table 3, or a wild-type Zika virus.

[0098] In some embodiments, the polynucleotide encoding a derivative of Zika virus prM / M comprises at least one substitution, at least one deletion, and / or at least one insertion compared to the wild-type Zika virus prM / M. In some embodiments, the Zika virus prM / M or a derivative of Zika virus prM / M comprises a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1F, or Table 1K. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus prM / M that encodes an amino acid sequence that is at least 95% identical to the wild-type prM / M sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus prM / M that encodes an amino acid sequence that is at least 96% identical to the wild-type prM / M sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus prM / M that encodes an amino acid sequence that is at least 97% identical to the wild-type prM / M sequence. In certain embodiments, the nucleic acid compositions comprise derivatives of Zika virus prM / M that encode an amino acid sequence that is at least 98% identical to the wild-type prM / M sequence. In certain embodiments, the nucleic acid compositions comprise derivatives of Zika virus prM / M that encode an amino acid sequence that is at least 99% identical to the wild-type prM / M sequence.

[0099] In some embodiments, the polynucleotide encoding a derivative of Zika virus E comprises at least one substitution, at least one deletion, and / or at least one insertion compared to wild-type Zika virus E. In some embodiments, the polynucleotide encoding E is translated into wild-type Zika virus E. In some embodiments, the Zika virus E or derivative of Zika virus E comprises a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1E, or Table 1J. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus E that encodes an amino acid sequence that is at least 95% identical to the wild-type E sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus E that encodes an amino acid sequence that is at least 96% identical to the wild-type E sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus E that encodes an amino acid sequence that is at least 97% identical to the wild-type E sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus E that encodes an amino acid sequence that is at least 98% identical to the wild-type E sequence. In certain embodiments, the nucleic acid composition comprises a derivative of Zika virus E that encodes an amino acid sequence that is at least 99% identical to the wild-type E sequence.

[0100] In some embodiments, the nucleic acid composition comprises a 5' untranslated region (5'UTR) of a Zika virus. In some embodiments, the nucleic acid composition comprises a 3' untranslated region (3'UTR) of a Zika virus. Non-limiting examples of untranslated sequences are shown in Table 1A, Table IB, Table 1C, Table 1G, Table 1H, or Table 2. In some embodiments, the Zika virus 5'UTR comprises a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1C, Table 1H, or Table 2. In some embodiments, the Zika virus 3'UTR comprises a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table IB, Table 1G, or Table 2.

[0101] In some embodiments, the nucleic acid composition does not include a polynucleotide encoding one or more nonstructural (NS) proteins selected from (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) two or more of (i)-(vii). For example, if the nucleic acid does not include such a polynucleotide, there is additional space in the composition to accommodate a large exogenous polynucleotide (e.g., up to about 10,000 or 6,000 bases).

[0102] In other embodiments, the nucleic acid composition comprises a polynucleotide encoding one or more nonstructural (NS) proteins selected from two or more of: (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii)(i)-(vii).

[0103] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding an NS1 protein. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding an NS2A protein. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding an NS2B protein. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding an NS3 protein. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding an NS4A protein. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding an NS4B protein. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding an NS5 protein.

[0104] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1 and NS2A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1 and NS2B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1 and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1 and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1 and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1 and NS5.

[0105] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A and NS2B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A and NS5.

[0106] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2B and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2B and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2B and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2B and NS5.

[0107] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS3 and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS3 and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS3 and NS5.

[0108] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS4A and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS4A and NS5.

[0109] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS4B and NS5.

[0110] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2A, and NS2B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2A, and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2A, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2A, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2A, and NS5.

[0111] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2B, and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2B, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2B, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS2B, and NS5.

[0112] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS3, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS3, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS3, and NS5.

[0113] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS4A, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS1, NS4A, and NS5.

[0114] In certain embodiments, the nucleic acid composition comprises polynucleotides encoding NS1, NS4B, and NS5.

[0115] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS2B, and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS2B, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS2B, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS2B, and NS5.

[0116] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS3, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS3, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS3, and NS5.

[0117] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS4A, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS4A, and NS5.

[0118] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2A, NS4B, and NS5.

[0119] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2B, NS3, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2B, NS3, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2B, NS3, and NS5.

[0120] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS2B, NS4A, and NS5.

[0121] In certain embodiments, the nucleic acid composition comprises polynucleotides encoding NS2B, NS4B, and NS5.

[0122] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS3 and NS4A and NS5.

[0123] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS3, NS4B, and NS5.

[0124] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding NS4A, NS4B, and NS5.

[0125] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2A, and NS2B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2A, and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2A, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2A, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2A, and NS5.

[0126] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2B, and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2B, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2B, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS2B, and NS5.

[0127] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS3, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS3, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1, NS3, and NS5.

[0128] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS1, NS4A, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS1, NS4A, and NS5.

[0129] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS1, NS4B, and NS5.

[0130] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS2B, and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS2B, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS2B, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS2B, and NS5.

[0131] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS3, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS3, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS3, and NS5.

[0132] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS4A, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A, NS4A, and NS5.

[0133] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS2A, NS4B, and NS5.

[0134] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2B, NS3, and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2B, NS3, and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2B, NS3, and NS5.

[0135] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS2B, NS4A, and NS5.

[0136] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS2B, NS4B, and NS5.

[0137] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS3 and NS4A, and NS5.

[0138] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS3, NS4B, and NS5.

[0139] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS4A, NS4B, and NS5.

[0140] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1 and NS2A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1 and NS2B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1 and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1 and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1 and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1 and NS5.

[0141] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A and NS2B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A and NS5.

[0142] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2B and NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2B and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2B and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2B and NS5.

[0143] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS3 and NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS3 and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS3 and NS5.

[0144] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS4A and NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS4A and NS5.

[0145] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except for NS4B and NS5.

[0146] In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS1. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS2B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS3. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS4A. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS4B. In certain embodiments, the nucleic acid composition comprises a polynucleotide encoding any NS protein except NS5.

[0147] In some embodiments, NS1 is Zika virus NS1. In some embodiments, NS2A is Zika virus NS2A. In some embodiments, NS2B is Zika virus NS2B. In some embodiments, NS3 is Zika virus NS3. In some embodiments, NS4A is Zika virus NS4A. In some embodiments, NS4B is Zika virus NS4B. In some embodiments, NS5 is Zika virus NS5.

[0148] In some embodiments, the nucleic acid composition comprises a derivative of NS1 comprising an amino acid sequence that is at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to the wild-type NS1 sequence of Table 1A. In some embodiments, the nucleic acid composition comprises a derivative of NS2A comprising an amino acid sequence that is at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to the wild-type NS2A sequence of Table 1A. In some embodiments, the nucleic acid composition comprises a derivative of NS2B comprising an amino acid sequence that is at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to the wild-type NS2B sequence of Table 1A. In some embodiments, the nucleic acid composition comprises a derivative of NS3 comprising 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 the wild-type NS3 sequence of Table 1A. In some embodiments, the nucleic acid composition comprises a derivative of NS4A comprising 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 the wild-type NS4A sequence of Table 1A. In some embodiments, the nucleic acid composition comprises a derivative of NS4B comprising 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 the wild-type NS4B sequence of Table 1A. In some embodiments, the nucleic acid composition comprises a derivative of NS5 comprising 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 the wild-type NS5 sequence.

[0149] In some embodiments, the components of the nucleic acid composition include African Zika virus components, Asian Zika virus components, or Brazilian Zika virus components, or combinations thereof. In one particular embodiment, the Zika virus is the African MR766 strain.

[0150] Exogenous Polynucleotides In some embodiments, the exogenous polynucleotide is about 10000, 9000, 8000, 7000, 6000, or 5000, or up to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length. For example, the exogenous polynucleotide is about 50 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length. The exogenous polynucleotide may be about 100 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length. The exogenous polynucleotide may be about 50 to about 6000 nucleotides in length. The exogenous polynucleotide may be about 100 to about 6000 nucleotides in length. The exogenous polynucleotide may be about 50 to about 5000 nucleotides in length. The exogenous polynucleotide may be from about 100 to about 5000 nucleotides in length.

[0151] In some embodiments, the exogenous polynucleotide is about 0.1 kb nucleotides in length. In other embodiments, the exogenous sequence is about 0.2 kb in length. In other embodiments, the exogenous sequence is about 0.3 kb in length. In other embodiments, the exogenous sequence is about 0.4 kb in length. In other embodiments, the exogenous sequence is about 0.5 kb in length. In other embodiments, the exogenous sequence is about 0.6 kb in length. In other embodiments, the exogenous sequence is about 0.7 kb in length. In other embodiments, the exogenous sequence is about 0.8 kb in length. In other embodiments, the exogenous sequence is about 0.9 kb in length. In other embodiments, the exogenous sequence is about 1 kb nucleotides in length. In other embodiments, the exogenous polynucleotide is about 2 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 3 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 4 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 5 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 6 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 7 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 8 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 9 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 10 kb nucleotides in length.

[0152] In some embodiments, the exogenous polynucleotide is an antigen or an antigenic epitope thereof. In some embodiments, the antigen or an antigenic epitope thereof is disease associated. In certain embodiments, the antigen or an antigenic epitope thereof is derived from a pathogen.

[0153] In some embodiments, the pathogen is a virus. In certain embodiments, the virus is human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papilloma virus (HPV-6, HPV-11), measles virus, rabies virus, polio virus, or yellow fever virus.

[0154] In some embodiments, the pathogen is a bacteria. In certain embodiments, the bacteria is selected from the group consisting of Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, and the like. 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans.

[0155] In some embodiments, the pathogen is a fungus, an amoeba, or a parasite. In certain embodiments, the fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 These are Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0156] In some embodiments, the exogenous polynucleotide encodes a gene editing tool. In certain embodiments, the gene editing tool is selected from the group consisting of meganuclease-related agents, CRISPR-related agents, TALEN-related agents, and zinc finger-related agents.

[0157] In some embodiments, the exogenous polynucleotide is a small interfering RNA (siRNA), an antisense RNA, a microRNA (miRNA), a small or short hairpin RNA (shRNA), a guide RNA (gRNA), a clustered regularly interspaced short palindromic repeat RNA (crRNA), a trans-activating clustered regularly interspaced short palindromic repeat RNA (tracrRNA), an immunostimulatory oligonucleotide, an antisense nucleic acid, or a ribozyme. In certain embodiments, the exogenous polynucleotide targets beta-secretase 1 (BACE1) and / or amyloid precursor protein (APP).

[0158] In some embodiments, the exogenous polynucleotide encodes a polypeptide associated with a genetic disorder. In certain embodiments, the polypeptide is brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neprilysin inhibitor (NEP), endothelin-converting enzyme (ECE), cathepsin B (CTSB), apolipoprotein E2 (APOE2), SH3 and multiple ankyrin repeat domains protein (SHANK), neurturin (NRTN), glial cell line-derived neurotrophic factor (GDNF), brain dopamine neurotrophic factor (CDNF), vascular endothelial growth factor A (VEGF-A), or aromatic L-amino acid decarboxylase (AADC).

[0159] In some embodiments, the exogenous polynucleotide encodes a therapeutic or diagnostic agent. In certain embodiments, the therapeutic agent is an antibody-based therapeutic agent, a hormone, a cytokine, an inhibitor or antagonist of immune checkpoint regulators, an immune stimulatory molecule, or an agonist of immune costimulatory molecules. In further embodiments, the inhibitor or antagonist of immune checkpoint regulators is an anti-PD1 antibody. In another embodiment, 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 yet other embodiments, the cytokine is a lymphokine, a monokine, a polypeptide hormone, a growth hormone, a parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, a glycoprotein hormone, a follicle stimulating hormone (FSH), a thyroid stimulating hormone (TSH), a luteinizing hormone (LH), a hepatocyte growth factor, a fibroblast growth factor, a prolactin, a placental lactogen, a tumor necrosis factor-α and -β, a Mullerian inhibitory substance, a mouse gonadotropin-related peptide, an inhibin, an activin, a vascular endothelial growth factor, an integrin, a thrombopoietin (TPO), These include nerve growth factor, NGF-β, platelet growth factor, transforming growth factor (TGF), TGF-α, TGF-β, insulin-like growth factor-I and -II, erythropoietin (EPO), osteoinductive factor, interferon, e.g., interferon-α, -β, and -γ, colony stimulating factor (CSF), macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF), granulocyte CSF (GCSF), interleukin (IL), tumor necrosis factor, TNF-α, TNF-β, LIF, or Kit ligand (KL), or a combination of two or more thereof.

[0160] Further feature examples In some embodiments, the components of the nucleic acid composition are expressed on one nucleic acid or two or more separate nucleic acids. In some embodiments, the nucleic acid composition further comprises one or more expression control elements in operable linkage that cause the expression of the nucleic acid composition in vitro or in vivo. In some embodiments, the expression control element is a promoter that drives the expression of the nucleic acid complex in vitro. In certain embodiments, the promoter is T7, T3, SP6, or any phage promoter.

[0161] 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 a CMV, SV40, or any eukaryotic promoter.

[0162] In some embodiments, the target cell is a neuronal cell, or a non-neuronal cell, VERO, COS, CHO, C6 / 36, HeLa, HEK, or HepG2. In some embodiments, the target cell is an oligodendrocyte, a microglia, or an astrocyte.

[0163] In some embodiments, provided herein are pharmaceutical compositions comprising the above-described nucleic acid compositions, and pharma- ceutically acceptable salts or derivatives thereof.

[0164] Recombinant Zika virus or Zika virus-like particles Viral vectors are widely used in fields such as gene therapy.Currently, three main vectors are based on adenovirus, adeno-associated virus, and lentivirus.However, each vector has limitations.Provided herein is a recombinant Zika virus or Zika virus-like particle, which in some embodiments provides flexible encapsulation strategy, similar or improved delivery efficiency, reduced insertion mutagenesis, reduced immune response, safety, or special targeting, or any combination thereof.

[0165] In some embodiments, recombinant Zika virus or Zika virus-like particles are generated from expressing a nucleic acid composition described herein in a producer cell that is further infected with a second Zika virus, such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0166] In certain embodiments, the recombinant Zika virus or Zika virus-like particle is generated from a nucleic acid composition that includes polynucleotides encoding only Zika virus C or a derivative of Zika virus C. In further specific embodiments, the recombinant Zika virus or Zika virus-like particle is generated from a nucleic acid composition that does not encode Zika virus C or a derivative of Zika virus C, does not encode Zika virus prM / M or a derivative of Zika virus prM / M, does not encode Zika virus E or a derivative of Zika virus E, or any combination of two or more of these. Thus, in some embodiments, the recombinant Zika virus described herein is generated from expressing such a nucleic acid composition in a producer cell that is further infected with a second Zika virus, such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0167] In certain embodiments, recombinant Zika virus or Zika virus-like particles are generated from nucleic acid compositions that include polynucleotides encoding only Zika virus prM / M or a derivative of Zika virus prM / M. Thus, in some embodiments, recombinant Zika viruses described herein are generated from expressing such nucleic acid compositions in a producer cell that is further infected with a second Zika virus, such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0168] In certain embodiments, recombinant Zika virus or Zika virus-like particles are generated from nucleic acid compositions that include polynucleotides encoding only Zika virus E or a derivative of Zika virus E. Thus, in some embodiments, recombinant Zika viruses described herein are generated from expressing such nucleic acid compositions in a producer cell that is further infected with a second Zika virus such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0169] In certain embodiments, recombinant Zika virus or Zika virus-like particles are generated from nucleic acid compositions that include a polynucleotide encoding Zika virus C or a derivative of Zika virus C, and a polynucleotide encoding only Zika virus prM / M or a derivative of Zika virus prM / M. Thus, in some embodiments, recombinant Zika viruses described herein are generated from expressing such nucleic acid compositions in a producer cell that is further infected with a second Zika virus, such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0170] In certain embodiments, the recombinant Zika virus or Zika virus-like particle is generated from a nucleic acid composition that includes a polynucleotide encoding Zika virus C or a derivative of Zika virus C, and a polynucleotide encoding only Zika virus E or a derivative of Zika virus E. Thus, in some embodiments, the recombinant Zika viruses described herein are generated from expressing such a nucleic acid composition in a producer cell that is further infected with a second Zika virus, such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0171] In certain embodiments, recombinant Zika virus or Zika virus-like particles are generated from nucleic acid compositions that include a polynucleotide encoding Zika virus prM / M or a derivative of Zika virus prM / M, and a polynucleotide encoding only Zika virus E or a derivative of Zika virus E. Thus, in some embodiments, recombinant Zika viruses described herein are generated from expressing such nucleic acid compositions in a producer cell that is further infected with a second Zika virus, such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0172] In some embodiments, the second Zika virus used in further infection is a wild type Zika virus. In some embodiments, the wild type Zika virus is an African strain, an Asian strain, and a Brazilian strain. In other embodiments, the second Zika virus used in further infection is a modified Zika virus. In some specific embodiments, the modified Zika virus comprises one or more microRNA-based gene silencing mechanisms. In some specific embodiments, the one or more microRNA-based gene silencing mechanisms control viral replication.

[0173] In some embodiments, the recombinant Zika virus or Zika virus-like particle is generated from expressing a nucleic acid described herein in a producer cell that is not infected with a second Zika virus, such that Zika virus C or a derivative of Zika virus C, Zika virus prM / M or a derivative of Zika virus prM / M, and Zika virus E or a derivative of Zika virus E are present in the recombinant Zika virus or Zika virus-like particle.

[0174] In some embodiments, producer cells are Vero E6, HEK, HEK 293T, HEK 293TT, FreeStyle™ 293-F cells, HEK-293.2sus, or C6 / 36 cells.

[0175] In some embodiments, the recombinant Zika virus is replication competent. In other embodiments, the recombinant Zika virus is unable to replicate without reducing vector titer or compromising expression of the exogenous polynucleotide. In some embodiments, the recombinant Zika virus or Zika virus-like particle has reduced insertional mutagenesis. In some embodiments, the recombinant Zika virus or Zika virus-like particle has a reduced immune response. In some embodiments, the Zika virus or Zika virus-like particle has tropism for cells of the central nervous system.

[0176] Methods for Recombinant Zika Virus or Zika Virus-Like Particles Comprising Three Structural Zika Proteins In another aspect, provided herein is a method of delivering an exogenous polynucleotide to a target cell, the method comprising applying to the target cell a recombinant Zika virus or Zika virus-like particle described herein comprising an exogenous polynucleotide.

[0177] In some embodiments, the target cell is a neuronal cell or a non-neuronal cell. In certain embodiments, the neuronal cell is an oligodendrocyte, a microglia, or an astrocyte. In another specific embodiment, the non-neuronal cell is a prostate epithelial cell, a urethral epithelial cell, a Sertoli cell, a Leydig cell, a spermatogonium cell, or a retinal cell.

[0178] In some embodiments, the methods are performed in vitro, ex vivo, or in vivo.

[0179] In some embodiments, the target cells express the exogenous polynucleotide transiently after delivery, hi other embodiments, the target cells express the exogenous polynucleotide persistently after delivery.

[0180] In another aspect, provided herein is a method of treating Alzheimer's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding brain-derived neurotrophic factor (BDNF).

[0181] In another aspect, provided herein is a method of treating Alzheimer's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide targeting amyloid precursor protein beta site cleaving enzyme 1 (BACE1).

[0182] In another aspect, provided herein is a method of treating Alzheimer's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide that targets amyloid precursor protein (APP).

[0183] In another aspect, provided herein is a method of treating Alzheimer's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a tau-targeting polynucleotide.

[0184] In another aspect, provided herein is a method of treating autism in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein, the recombinant Zika virus or Zika virus-like particle comprising a polynucleotide encoding an SH3 and multiple ankyrin repeat domains protein (SHANK).

[0185] In yet another aspect, provided herein is a method of treating Parkinson's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding aromatic L-amino acid decarboxylase (AADC).

[0186] In another aspect, provided herein is a method of treating Parkinson's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding human aromatic L-amino acid decarboxylase.

[0187] In another aspect, provided herein is a method of treating Parkinson's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding a glial cell line-derived neurotrophic factor.

[0188] In another aspect, provided herein is a method of treating / managing Down's syndrome in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide that targets further replication of HSA21.

[0189] In another aspect, provided herein is a method of treating choroideremia-blindness in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding Rab escort protein 1 (REP1).

[0190] In another aspect, provided herein is a method of treating Leber congenital amaurosis in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding RPE65.

[0191] In another aspect, provided herein is a method of treating Parkinson's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein, the recombinant Zika virus or Zika virus-like particle comprising a polynucleotide encoding glutamic acid decarboxylase.

[0192] In another aspect, provided herein is a method of treating ornithine transcarbamylase (OTC) in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding ornithine transcarbamylase.

[0193] In another aspect, provided herein is a method of treating multiple sclerosis in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding interferon beta-1a and 1b.

[0194] In another aspect, provided herein is a method of treating Pompe disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding an acid alpha-glucosidase.

[0195] In another aspect, provided herein is a method of treating depression in a subject in need thereof, the method comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described herein comprising a polynucleotide encoding a selective serotonin reuptake inhibitor.

[0196] In some embodiments, the administering is by systemic delivery.

[0197] In some embodiments, the administering step is performed intravenously and / or intratumorally.

[0198] In some embodiments, the administering step targets ventricular, cisternal, subpial, and / or intrathecal cerebrospinal fluid, hi other embodiments, the administering step is not performed intraparenchymal.

[0199] Medication and Treatment Regimens The frequency of administration of pharmaceutical compositions containing a recombinant Zika virus or Zika virus-like particle provided herein can vary based on the method performed, the physical characteristics of the subject, the severity of the cancer, the type of cancer, and the formulation and means used to administer the composition.

[0200] The duration of treatment may be determined by the attending physician based on the disease being treated. The duration of administration often varies depending on a number of factors. Exemplary factors include, but are not limited to, the patient's response, the severity of symptoms, and the type of cancer. Under some conditions, treatment continues for days, weeks, or months. Under other conditions, complete treatment is achieved by administering one, two, or three doses of the pharmaceutical composition over the entire course of treatment. In certain embodiments, complete treatment may be achieved using a single dose of the pharmaceutical composition.

[0201] In certain embodiments where a patient's condition improves, the dose of a recombinant Zika virus or Zika virus-like particle described herein, or a pharmaceutical composition thereof, administered may be temporarily reduced or suspended (i.e., a "drug holiday") for a certain period of time.

[0202] In certain embodiments, the dose of the composition being administered is temporarily reduced or temporarily suspended (ie, "drug diversion") for a certain period of time.

[0203] In some embodiments, once improvement of the patient's disease has occurred, a maintenance dose is administered as needed. In certain embodiments, the dosage or frequency of administration, or both, is then reduced, depending on the symptoms, to a level at which improvement of the disease, disorder, or condition is maintained. However, in certain embodiments, the patient requires intermittent treatment on a long-term basis upon any recurrence of symptoms.

[0204] The amount of recombinant Zika virus or Zika virus-like particle provided herein will vary depending on factors such as the particular virus, the state and severity of the disease, the identity of the subject in need of treatment (e.g., weight, sex), etc., but may nevertheless be determined according to the particular circumstances surrounding the case. In some embodiments, the desired dose is conveniently provided in a single dose or in divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, e.g., as subdoses two, three, four or more times daily.

[0205] In some embodiments, administration of a recombinant Zika virus or Zika virus-like particle provided herein is according to potency, and the titer of the recombinant Zika virus or Zika virus-like particle is about 10 6 PFU / mL~about 10 10 PFU / mL.

[0206] In some embodiments, the amount of recombinant Zika virus or Zika virus-like particles of the present disclosure administered to a subject is about 10 3 ~about 10 12 It may be an infectious viral particle or a plaque forming unit (PFU).

[0207] In some embodiments, the recombinant Zika virus or Zika virus-like particle of the present disclosure comprises at least about 10 3 Virus particles / dose ~ approx. 10 14 It may be administered in a dose that may contain viral particles / dose.

[0208] In some embodiments, the recombinant Zika virus or Zika virus-like particle of the present disclosure comprises at least about 10 3 PFU / kg ~ approx. 10 14 It may be administered in a dose that may contain PFU / kg.

[0209] In some embodiments, the recombinant Zika virus or Zika virus-like particle of the present disclosure comprises at least about 10 3 Virus particles / kg ~ approx. 10 14 It may be administered in a dose that may contain viral particles / kg.

[0210] Pharmaceutical Compositions and Formulations of Recombinant Zika Virus or Zika Virus-Like Particles Provided herein are recombinant Zika viruses or Zika virus-like particles formulated in pharmaceutical compositions.

[0211] Pharmaceutical composition is formulated in a conventional manner, using one or more pharma- ceutical acceptable inactive ingredients, which facilitates the processing of active agent into pharma- ceutical preparations.The summary of pharmaceutical composition described herein can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins, 1999), and such disclosure is incorporated herein by reference. For some virus delivery methods, a pharma- ceutically acceptable vehicle is selected from known pharma- ceutically acceptable vehicles for delivery and should be one in which the virus is stable.

[0212] Further provided herein are pharmaceutical compositions comprising a virus, a pharma- ceutically acceptable inactive ingredient, another drug or pharmaceutical agent, a carrier, an adjuvant, a preservative, a stabilizer, a wetting agent, or an emulsifier, a solution promoter, a salt, a buffer, an excipient, a binder, a filler, a suspending agent, a flavoring agent, a sweetening agent, a disintegration agent, a dispersing agent, a surfactant, a lubricant, a coloring agent, a diluent, a solubilizing agent, a wetting agent, a plasticizer, a permeation enhancer, an antifoaming agent, an antioxidant, a preservative, or a combination thereof.

[0213] Another aspect of the present disclosure provides a pharmaceutical composition comprising a recombinant Zika virus as described herein. In some embodiments, the pharmaceutical composition may comprise a solubilizer and an excipient. In some embodiments, the excipient may comprise one or more of a buffer, a stabilizer, an antioxidant, a binder, a diluent, a dispersant, a rate controlling agent, a lubricant, a glidant, a disintegrant, a plasticizer, a preservative, or any combination thereof. In some embodiments, the excipient may comprise sodium dihydrogen phosphate dihydrate, sodium hydrogen phosphate dihydrate, sodium chloride, myo-inositol, sorbitol, or any combination thereof. In some embodiments, the pharmaceutical composition does not comprise a preservative. In some embodiments, the pharmaceutical composition may comprise one or more of a preservative, a diluent, and a carrier. In some embodiments, the pharmaceutical composition may comprise an additional active ingredient or a salt thereof. In some embodiments, the solubilizer may be sterile water. In some embodiments, the pharmaceutical composition may comprise an additional active ingredient, where the additional active ingredient may be an additional oncolytic virus.

[0214] Another aspect of the present disclosure provides a method of enhancing the therapeutic effect of an oncolytic virus when the virus is delivered systemically to a subject, the method comprising systemic administration of a recombinant Zika virus or Zika virus-like particle disclosed herein, a recombinant Zika virus or Zika virus-like particle described herein, or a pharmaceutical composition disclosed herein.

[0215] The pharmaceutical compositions containing the modified virus, which is a recombinant Zika virus or Zika virus-like particle described herein, may be prepared as a solution, a dispersion in glycerol, a liquid polyethylene glycol, in oil, in a solid dosage form, as an inhalable dosage form, as an intranasal dosage form, as a liposomal formulation, as a dosage form containing nanoparticles, a dosage form containing microparticles, a polymeric dosage form, or any combination thereof. In some embodiments, the pharmaceutical compositions described herein may include a stabilizer and a buffering agent. In some embodiments, the pharmaceutical compositions described herein may include a solubilizing agent, such as sterile water, Tris buffer, etc. In some embodiments, the pharmaceutical compositions described herein may include an excipient. The excipient may be an excipient described in the Handbook of Pharmaceutical Excipients, American Pharmaceutical Association (1986). Non-limiting examples of suitable excipients may include buffering agents, preservatives, stabilizers, binders, compression agents, lubricants, chelating agents, dispersion enhancers, disintegrants, flavoring agents, sweetening agents, and coloring agents.

[0216] In some embodiments, the excipient may be a buffering agent. In some embodiments, the excipient may include a preservative. Non-limiting examples of suitable preservatives may include antioxidants and antimicrobial agents. In some embodiments, the pharmaceutical compositions described herein may include a binder as an excipient. In some embodiments, the pharmaceutical compositions described herein may include a lubricant as an excipient. In some embodiments, the pharmaceutical formulation may include a dispersion enhancer as an excipient. In some embodiments, the pharmaceutical compositions described herein may include a disintegrant as an excipient. In some cases, the pharmaceutical compositions described herein may include a chelating agent.

[0217] Also contemplated are combination products comprising one or more recombinant Zika viruses or one or more Zika virus-like particles described herein.

[0218] Under normal storage and use conditions, the pharmaceutical compositions described herein may contain preservatives to prevent the growth of microorganisms.In certain cases, the pharmaceutical compositions described herein may be free of preservatives.The pharmaceutical forms suitable for use by injection may include sterile aqueous solutions or dispersions, and sterile powders for extemporaneous preparation of sterile injectable solutions or dispersions.Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents.

[0219] For parenteral administration in aqueous solution, for example, the liquid dosage form may be suitably buffered if necessary, and the liquid diluent may be made isotonic with sufficient saline or glucose. The liquid dosage form is particularly suitable for intravenous, intramuscular, subcutaneous, intratumoral, and intraperitoneal administration. In this regard, usable sterile aqueous media will be known to those skilled in the art in light of the present disclosure. For example, a dose may be dissolved in 1 mL to 20 mL of isotonic NaCl solution and added to 100 mL to 1000 mL of fluid, for example, sodium bicarbonate buffered saline, or injected into the proposed injection site.

[0220] In certain embodiments, a sterile injectable solution may be prepared by incorporating a recombinant Zika virus or Zika virus-like particle according to the present disclosure, a recombinant Zika virus or Zika virus-like particle described herein, or a pharmaceutical composition containing the recombinant Zika virus or Zika virus-like particle in the required amount in an appropriate solvent with various other ingredients as listed above, as required, followed by filtered sterilization. In general, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle containing the basic dispersion medium and the required other ingredients from those listed above. The compositions disclosed herein may be formulated in neutral or salt forms. Upon formulation, the pharmaceutical compositions may be administered in a manner compatible with the dosage formulation and in a therapeutically effective amount.

[0221] In certain embodiments, the pharmaceutical composition of the present disclosure may comprise an effective amount of the recombinant virus disclosed herein in combination with a pharma- ceutically acceptable carrier. As used herein, "pharmaceutically acceptable" includes any carrier that does not interfere with the effectiveness of the biological activity of the active ingredient and / or is not toxic to the patient to whom the carrier is administered. Non-limiting examples of suitable pharmaceutical carriers include phosphate buffered saline solutions, water, emulsions, such as oil / water emulsions, various types of wetting agents, and sterile solutions. Further non-limiting examples of pharma- ceutically compatible carriers may include gels, bioadsorbent matrix materials, placement elements containing recombinant Zika virus or Zika virus-like particles, or any other suitable vehicle, delivery, or administration means or material. Such carriers may be formulated by conventional methods and administered to a subject in an effective amount.

[0222] Recombinant Zika virus or Zika virus-like particle kits In one aspect of the disclosure, a kit is provided herein that includes one or more reagents or devices for carrying out the methods disclosed herein. In some embodiments, the kit includes a recombinant Zika virus or Zika virus-like particle provided herein. In some embodiments, the kit includes a means for administering the recombinant Zika virus or Zika virus-like particle provided herein.

[0223] In some embodiments, the kit includes suitable instructions for carrying out the method of the kit. The instructions may provide information for carrying out all of the methods disclosed herein, regardless of whether the method is carried out using only the reagents provided in the kit. The kit and instructions may require additional reagents or systems.

[0224] Kits and articles of manufacture are also described herein for use in the therapeutic applications described herein. In some embodiments, such kits include carriers, packages, or containers that are compartmentalized to receive one or more containers, such as vials, tubes, each of which contains one of the separate elements used in the methods described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers may be formed from a variety of materials, such as glass or plastic. The articles of manufacture provided herein include 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 the formulation selected and its intended mode of administration and treatment. The containers optionally have a sterile access port (e.g., the container is an intravenous solution bag or vial with a stopper that can be pierced by a hypodermic needle). Such kits optionally include the composition and an identifying description or label or instructions for its use in the methods described herein.

[0225] The kit typically includes one or more additional containers, each containing one or more of a variety of materials (e.g., reagents, optionally in concentrated form, and / or devices) desirable from a commercial or user standpoint for use of the recombinant Zika virus or Zika virus-like particles described herein. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes, carriers, packaging, container, vial and / or tube labels listing contents and / or instructions for use, and inserts containing instructions for use. A set of instructions is also typically included.

[0226] In some embodiments, the label is on or associated with the container. A label may be on a container when letters, numbers, or other characters forming the label are attached, molded, or etched onto the container itself, and a label may be associated with a container when present within a receptacle or carrier that also holds the container, e.g., as a package insert. A label may be used to indicate that the contents are to be used for a particular therapeutic application. A label may also indicate instructions for the use of the contents, such as in the methods described herein.

[0227] In certain embodiments, pharmaceutical compositions comprising recombinant Zika virus or Zika virus-like particles provided herein and optional additional active agents are provided in a pack or dispenser device that can contain one or more unit dosage forms. The pack may, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied by a notice associated with the container in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, which notice reflects approval by the agency of the drug form for human or veterinary administration. Such notice may, for example, be a label approved for prescription drugs by the US Food and Drug Administration, or an approved package insert. Compositions containing recombinant Zika virus or Zika virus-like particles described herein formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated disease.

[0228] Nucleic Acid Compositions Comprising Regulatory Regions of the Zika Genome The wild-type Zika genome contains several regulatory regions, including the 5'UTR and 3'UTR. The UTRs have been shown to be important for viral replication and immune regulation. Elements in the UTRs have been shown to be essential for genome circularization, which initiates RNA synthesis.

[0229] Ribozymes are self-cleaving RNAs. Small ribozyme motifs are mainly classified into four types: hammerhead, hairpin, Varkud satellite (VS), and hepatitis delta virus (HDV).

[0230] In some embodiments, a nucleic acid composition described herein comprises a polynucleotide encoding a ribozyme. In some embodiments, a Zika virus or Zika virus-like particle described herein comprises a ribozyme.

[0231] In one aspect, provided herein is a nucleic acid composition comprising (i) a polynucleotide encoding a ribozyme, (ii) a 5' untranslated region (UTR) of a Zika virus, (iii) a 3' UTR of a Zika virus, and (iv) a polynucleotide exogenous to a Zika virus. In some embodiments, the ribozyme is a hammerhead ribozyme or a 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 D Virus (HDV) ribozyme. In some embodiments, the polynucleotide encoding the ribozyme encodes a hammerhead ribozyme and the polynucleotide encoding the second ribozyme encodes a HDV ribozyme. In some embodiments, the nucleic acid composition further comprises a C moiety.

[0232] In some embodiments, the Zika virus is an African Zika virus, an Asian Zika virus, a component of a Brazilian Zika virus, or a combination of one or two of these. In certain embodiments, the Zika virus is the African MR766 strain.

[0233] Exogenous Polynucleotides In some embodiments, the exogenous polynucleotide is an antigen or an antigenic epitope thereof. In some embodiments, the antigen or an antigenic epitope thereof is disease associated. In certain embodiments, the antigen or an antigenic epitope thereof is derived from a pathogen.

[0234] In some embodiments, the pathogen is a virus. In certain embodiments, the virus is human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papilloma virus (HPV-6, HPV-11), measles virus, rabies virus, polio virus, or yellow fever virus.

[0235] In some embodiments, the pathogen is a bacteria. In certain embodiments, the bacteria is selected from the group consisting of Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, and the like. 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans.

[0236] In some embodiments, the pathogen is a fungus, an amoeba, or a parasite. In certain embodiments, the fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 These are Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0237] In some embodiments, the exogenous polynucleotide encodes a gene editing tool. In certain embodiments, the gene editing tool is selected from the group consisting of meganuclease-related agents, CRISPR-related agents, TALEN-related agents, and zinc finger-related agents.

[0238] In some embodiments, the exogenous polynucleotide is a small interfering RNA (siRNA), an antisense RNA, a microRNA (miRNA), a small or short hairpin RNA (shRNA), a guide RNA (gRNA), a clustered regularly interspaced short palindromic repeat RNA (crRNA), a trans-activating clustered regularly interspaced short palindromic repeat RNA (tracrRNA), an immunostimulatory oligonucleotide, an antisense nucleic acid, or a ribozyme. In certain embodiments, the exogenous polynucleotide targets beta-secretase 1 (BACE1) and / or amyloid precursor protein (APP).

[0239] In some embodiments, the exogenous polynucleotide encodes a polypeptide associated with a genetic disorder. In certain embodiments, the polypeptide is brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neprilysin inhibitor (NEP), endothelin-converting enzyme (ECE), cathepsin B (CTSB), apolipoprotein E2 (APOE2), SH3 and multiple ankyrin repeat domains protein (SHANK), neurturin (NRTN), glial cell line-derived neurotrophic factor (GDNF), brain dopamine neurotrophic factor (CDNF), vascular endothelial growth factor A (VEGF-A), or aromatic L-amino acid decarboxylase (AADC).

[0240] In some embodiments, the exogenous polynucleotide encodes a therapeutic or diagnostic agent. In certain embodiments, the therapeutic agent is an antibody-based therapeutic agent, a hormone, a cytokine, an inhibitor or antagonist of immune checkpoint regulators, an immune stimulatory molecule, or an agonist of immune costimulatory molecules. In further embodiments, the inhibitor or antagonist of immune checkpoint regulators is an anti-PD1 antibody. In another embodiment, 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 yet other embodiments, the cytokine is a lymphokine, a monokine, a polypeptide hormone, a growth hormone, a parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, a glycoprotein hormone, a follicle stimulating hormone (FSH), a thyroid stimulating hormone (TSH), a luteinizing hormone (LH), a hepatocyte growth factor, a fibroblast growth factor, a prolactin, a placental lactogen, a tumor necrosis factor-α and -β, a Mullerian inhibitory substance, a mouse gonadotropin-related peptide, an inhibin, an activin, a vascular endothelial growth factor, an integrin, a thrombopoietin (TPO), These include nerve growth factor, NGF-β, platelet growth factor, transforming growth factor (TGF), TGF-α, TGF-β, insulin-like growth factor-I and -II, erythropoietin (EPO), osteoinductive factor, interferon, e.g., interferon-α, -β, and -γ, colony stimulating factor (CSF), macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF), granulocyte CSF (GCSF), interleukin (IL), tumor necrosis factor, TNF-α, TNF-β, LIF, or Kit ligand (KL), or a combination of two or more thereof.

[0241] In some embodiments, the exogenous polynucleotide is about 10000, 9000, 8000, 7000, 6000, or 5000, or up to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length. For example, the exogenous polynucleotide is about 50 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length. The exogenous polynucleotide may be about 100 to about 10000, 9000, 8000, 7000, 6000, or 5000 nucleotides in length. The exogenous polynucleotide may be about 50 to about 6000 nucleotides in length. The exogenous polynucleotide may be about 100 to about 6000 nucleotides in length. The exogenous polynucleotide may be about 50 to about 5000 nucleotides in length. The exogenous polynucleotide may be from about 100 to about 5000 nucleotides in length.

[0242] In some embodiments, the exogenous polynucleotide is about 0.1 kb nucleotides in length. In other embodiments, the exogenous sequence is about 0.2 kb in length. In other embodiments, the exogenous sequence is about 0.3 kb in length. In other embodiments, the exogenous sequence is about 0.4 kb in length. In other embodiments, the exogenous sequence is about 0.5 kb in length. In other embodiments, the exogenous sequence is about 0.6 kb in length. In other embodiments, the exogenous sequence is about 0.7 kb in length. In other embodiments, the exogenous sequence is about 0.8 kb in length. In other embodiments, the exogenous sequence is about 0.9 kb in length. In other embodiments, the exogenous sequence is about 1 kb nucleotides in length. In other embodiments, the exogenous polynucleotide is about 2 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 3 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 4 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 5 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 6 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 7 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 8 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 9 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 10 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 11 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 12 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 13 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 14 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 15 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 16 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 17 kb nucleotides in length.In some embodiments, the exogenous polynucleotide is about 18 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 19 kb nucleotides in length. In some embodiments, the exogenous polynucleotide is about 20 kb nucleotides in length.

[0243] Other Properties of Nucleic Acid Compositions Comprising Regulatory Regions of the Zika Genome In some embodiments, the nucleic acid composition further comprises one or more expression control elements in operable linkage that cause 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 certain embodiments, the promoter is T7, T3, SP6, or any phage promoter.

[0244] In another aspect, provided herein is a pharmaceutical composition comprising a nucleic acid composition described herein, or a pharma- ceutically acceptable salt or derivative thereof.

[0245] Methods of Use of Nucleic Acid Compositions In another aspect, provided herein is a method of delivering an exogenous polynucleotide to a target cell, the method comprising administering to the target cell a compound generated from a nucleic acid composition described herein that includes the exogenous polynucleotide.

[0246] In some embodiments, the compound is produced by encapsulating the transcript produced from the nucleic acid composition described herein with a lipid-based agent. In certain embodiments, the transcript is uncapped. In another particular embodiment, the transcript is produced by transcribing the nucleic acid composition in vitro.

[0247] In some embodiments, the lipid-based agent is a lipofectamine-related reagent, a liposome, or a lipid nanoparticle.

[0248] In some embodiments, the lipid-based agents described herein are cationic lipids, neutral lipids, and polyethylene glycol conjugates, such as PEG-diacylglycerol, PEG-diacylglycamide, PEG-cholesterol, or PEG-DMB conjugates, cholesterol, or cholesterol derivatives.

[0249] Suitable cationic lipids include cationic lipids that carry a net negative charge at a selected pH, e.g., physiological pH. Particularly useful cationic lipids include cationic lipids that have a relatively small head group, e.g., a tertiary amine, a quaternary amine, or a guanidine head group, and an asymmetric lipid chain with steric hindrance. In any of the embodiments described herein, the cationic lipid may be N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (D OTMA), N,N-Dimethyl-2,3-dioleyloxy)propylamine (DODMA), 1,2-Dioleoyl-3-dimethylammonium-propane (DODAP), 1,2-Dioleoylcarbamyl-3-dimethylammonium-propane (DOCDAP), 1,2-Dilineoyl-3-dimethylammonium-propane (DLILDAP), Dioleoyloxy-N-[2-sperminecarboxamido)ethyl}-N,N-dimethyl Dimethylamino-1-propaneaminium trifluoroacetate (DOSPA), dioctadecylamidoglycylspermine (DOGS), DC-Chol, 1,2-dimyristyloxypropyl-3-dimethyl-hydroxyethylammonium bromide (DMRIE), 3-dimethylamino-2-(cholest-5-ene-3-β-oxybutane-4-oxy)-1-(cis,cis-9,12-octadecadienooxy)propane (CLinDMA), 2-[5'-(cholestane) Cationic lipids may be selected from the group consisting of cis-5-ene-3P-oxy)-3'-oxapentoxy)-3-dimethyl-1-(cis,cis-9',12'-octadecadienooxy)propane (CpLinDMA), N,N-dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N'-dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), and / or mixtures thereof, as well as other cationic lipids sharing similar properties.The above cationic lipids may include a variety of different salts known in the art.

[0250] In some embodiments, the head group of the cationic lipid can be attached to the lipid chain via a cleavable or non-cleavable linker, such as those described herein or otherwise known in the art. Non-limiting examples of suitable linkers include those that contain C1-C10 alkyl, alkyl ether, polyether, polyethylene glycol, acetal, amide, carbonyl, carbamide, carbamate, carbonate, ester (i.e., monoester, diester), or succinyl.

[0251] Suitable neutral lipids include any of a variety of neutral uncharged, zwitterionic, or anionic lipids that are capable of forming stable complexes, which are preferably neutral, but may be positively or negatively charged. In any of the embodiments described herein, suitable neutral lipids include those selected from the compound having the formula NLI-NLVII, dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), egg yolk phosphatidylcholine (EPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), -phosphatidylethanolamine (POPE) and dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), cholesterol, and other neutral lipids described herein below, and / or mixtures thereof.

[0252] Suitable polyethylene glycol-diacylglycerol or polyethylene glycol-diacylglycamide (PEGDAG) conjugates include those that contain a dialkylglycerol or dialkylglycamide group having an alkyl chain length that independently contains from about C4 to about C40 saturated or unsaturated carbon atoms. The dialkylglycerol or dialkylglycamide group may further contain one or more substituted alkyl groups. In any of the embodiments described herein, the PEG conjugate may be selected from PEG-dilaurylglycerol (C12), PEG-dimyristylglycerol (C14), PEG-dipalmitoylglycerol (C16), PEG-disterylglycerol (C18), PEG-dilaurylglycamide (C12), PEG-dimyristylglycamide (C14), PEG-dipalmitoylglycamide (C16), and PEG-disterylglycamide (C18), PEG-cholesterol (1-[8'-(cholest-5-ene-3P-oxy)carboxamido-3',6'-dioxaotanyl]carbamoyl-ω-methyl-poly(ethylene glycol), and PEG-DMB (3,4-ditetradeoxybenzyl-ω-methyl-poly(ethylene glycol) ether).

[0253] In another aspect, provided herein is a method of inducing or boosting an immune response in a subject, the method comprising administering to the subject an effective amount of a compound generated from a nucleic acid composition described herein that comprises an antigen or an epitope of the antigen.

[0254] In some embodiments, the compound is produced by encapsulating the transcript produced from the nucleic acid composition described herein with a lipid-based agent. In certain embodiments, the transcript is uncapped. In another particular embodiment, the transcript is produced by transcribing the nucleic acid composition in vitro.

[0255] In some embodiments, the lipid-based agent is a lipofectamine-related reagent, a liposome, or a lipid nanoparticle.

[0256] In some embodiments, the administering step is performed intramuscularly.

[0257] In some embodiments, the antigen or antigenic epitope thereof is derived from a pathogen.

[0258] In some embodiments, the pathogen is a virus. In certain embodiments, the virus is human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papilloma virus (HPV-6, HPV-11), measles virus, rabies virus, polio virus, or yellow fever virus.

[0259] In some embodiments, the pathogen is a bacteria. In certain embodiments, the bacteria is selected from the group consisting of Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, and the like. 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans.

[0260] In some embodiments, the pathogen is a fungus, an amoeba, or a parasite. In certain embodiments, the fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 These are Trypanosoma brucei, Trypanosoma cruzi, and Taenia multiceps.

[0261] Non-limiting example embodiments 1. A nucleic acid composition comprising: (i) a polynucleotide encoding a Zika virus 5'UTR or a derivative of a Zika virus 5'UTR, a polynucleotide encoding a Zika virus 3'UTR or a derivative of a 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 membrane protein (prM / M) or a derivative of a Zika virus prM / M, a polynucleotide encoding a Zika virus envelope protein (E) or a derivative of a Zika virus E, or any combination thereof; and (ii) a polynucleotide exogenous to a Zika virus. 2. The nucleic acid composition of embodiment 1, comprising a polynucleotide encoding a Zika virus 5'UTR, a polynucleotide encoding a Zika virus 3'UTR, a polynucleotide encoding a Zika virus C or a derivative of Zika virus C, a polynucleotide encoding a Zika virus prM / M or a derivative of Zika virus prM / M, and a polynucleotide encoding a Zika virus E or a derivative of Zika virus E. 3. The nucleic acid composition of embodiment 1 or embodiment 2, wherein the polynucleotide encoding the Zika virus 5'UTR or a derivative of the Zika virus 5'UTR, the polynucleotide encoding the Zika virus 3'UTR or a derivative of the Zika virus 3'UTR, the polynucleotide encoding the Zika virus C or a derivative of the Zika virus C, the polynucleotide encoding the Zika virus prM / M or a derivative of the Zika virus prM / M, and the polynucleotide encoding the Zika virus E or a derivative of the Zika virus E are expressed on one or more separate nucleic acids. 4. The nucleic acid composition of the preceding embodiment, 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 compared to a wild-type Zika virus 5'UTR. 5. The nucleic acid composition of the preceding embodiment, wherein the Zika virus 5'UTR or a derivative of the Zika virus 5'UTR comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1C, Table 1H, or Table 2. 6. The nucleic acid composition of the preceding embodiment, wherein the polynucleotide encoding a derivative of the Zika virus 3'UTR comprises at least one substitution, at least one deletion, and / or at least one insertion compared to a wild-type Zika virus 3'UTR. 7. The nucleic acid composition of the preceding embodiment, wherein the Zika virus 3'UTR or a derivative of the Zika virus 3'UTR comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table IB, Table 1G, or Table 2. 8. The nucleic acid composition of the preceding embodiment, wherein the polynucleotide encoding the derivative of Zika virus C comprises at least one substitution, at least one deletion, and / or at least one insertion compared to wild-type Zika virus C. 9. The nucleic acid composition of the preceding embodiment, wherein the Zika virus C or derivative of the Zika virus C comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table ID, Table 1I, Table 2, or Table 3. 10. The nucleic acid composition of the preceding embodiment, wherein the polynucleotide encoding the derivative of Zika virus prM / M comprises at least one substitution, at least one deletion, and / or at least one insertion compared to wild-type Zika virus prM / M. 11. The nucleic acid composition of the preceding embodiment, wherein the Zika virus prM / M or a derivative of the Zika virus prM / M comprises a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1F, or Table 1K. 12. The nucleic acid composition of the preceding embodiment, wherein the polynucleotide encoding the derivative of Zika virus E comprises at least one substitution, at least one deletion, and / or at least one insertion compared to wild-type Zika virus E. 13. The nucleic acid composition of the preceding embodiment, wherein the polynucleotide encoding E is translated into a wild-type Zika virus E. 14. The nucleic acid composition of the preceding embodiment, wherein the Zika virus E or derivative of Zika virus E comprises a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a sequence in Table 1A, Table 1E, or Table 1J. 15. The nucleic acid composition of the preceding embodiment, comprising the 5' untranslated region (5'UTR) of Zika virus. 16. The nucleic acid composition of the previous embodiment, comprising the 3' untranslated region (3'UTR) of Zika virus. 17. The nucleic acid composition of any preceding embodiment, which does not comprise a polynucleotide encoding one or more nonstructural (NS) proteins selected from two or more of (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) (i)-(vii). 18. The nucleic acid composition of any preceding embodiment, further comprising a polynucleotide encoding one or more nonstructural (NS) proteins selected from two or more of (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) (i)-(vii). 19. The nucleic acid composition of embodiment 17 or embodiment 18, wherein said NS1 is Zika virus NS1, said NS2A is Zika virus NS2A, said NS2B is Zika virus NS2B, said NS3 is Zika virus NS3, said NS4A is Zika virus NS4A, said NS4B is Zika virus NS4B, or said NS5 is Zika virus NS5, or any combination of two or more thereof. 20. The nucleic acid composition of the preceding embodiment, wherein the components of the nucleic acid composition include an African Zika virus component, an Asian Zika virus component, or a Brazilian Zika virus component, or a combination thereof. 21. The nucleic acid composition of embodiment 20, wherein the Zika virus is the African MR766 strain. 22. The nucleic acid composition of the previous embodiment, wherein the exogenous polynucleotide is an antigen or an antigenic epitope thereof. 23. The nucleic acid composition of embodiment 22, wherein the antigen or antigenic epitope thereof is disease associated. 24. The nucleic acid composition of embodiment 23, wherein the antigen or antigenic epitope thereof is derived from a pathogen. 25. The nucleic acid composition of embodiment 24, wherein the pathogen is a virus. 26. The nucleic acid composition of embodiment 25, wherein the virus is a human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papillomavirus (HPV-6, HPV-11), measles virus, rabies virus, poliovirus, or yellow fever virus. 27. The nucleic acid composition of embodiment 24, wherein the pathogen is a bacterium. 28. The bacterium is selected from the group consisting of Acinetobacter baumanii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp., Burkholderia cepacis, Burkholderia psedomallei, and Campylobacter jejuni. 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans. 29. The nucleic acid composition of embodiment 24, wherein the pathogen is a fungus, an amoeba, or a parasite. 30. The fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 brucei, Trypanosoma cruzi, and Taenia multiceps. 31. The nucleic acid composition of any one of embodiments 1 to 30, wherein the exogenous polynucleotide encodes a gene editing tool. 32. The nucleic acid composition of embodiment 31, wherein the gene editing tool is selected from the group consisting of meganuclease-related agents, CRISPR-related agents, TALEN-related agents, and zinc finger-related agents. 33. The nucleic acid composition according to any one of embodiments 1 to 30, wherein the exogenous polynucleotide is a small interfering RNA (siRNA), an antisense RNA, a microRNA (miRNA), a small or short hairpin RNA (shRNA), a guide RNA (gRNA), a clustered regularly interspaced short palindromic repeat RNA (crRNA), a trans-activating clustered regularly interspaced short palindromic repeat RNA (tracrRNA), an immunostimulatory oligonucleotide, an antisense nucleic acid, or a ribozyme. 34. The nucleic acid composition of embodiment 33, wherein the exogenous polynucleotide targets beta-secretase 1 (BACE1) and / or amyloid precursor protein (APP). 35. The nucleic acid composition of any one of embodiments 1 to 30, wherein the exogenous polynucleotide encodes a polypeptide associated with a genetic disorder. 36. The nucleic acid composition of embodiment 35, wherein the polypeptide is brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neprilysin inhibitor (NEP), endothelin-converting enzyme (ECE), cathepsin B (CTSB), apolipoprotein E2 (APOE2), SH3 and multiple ankyrin repeat domains protein (SHANK), neurturin (NRTN), glial cell line-derived neurotrophic factor (GDNF), brain dopamine neurotrophic factor (CDNF), vascular endothelial growth factor A (VEGF-A), or aromatic L-amino acid decarboxylase (AADC). 37. The nucleic acid composition of any one of embodiments 1 to 30, wherein the exogenous polynucleotide encodes a therapeutic or diagnostic agent. 38. The nucleic acid composition of embodiment 37, wherein 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 costimulatory molecule. 39. The nucleic acid composition of embodiment 38, wherein the inhibitor or antagonist of the immune checkpoint regulator is an anti-PD1 antibody. 40. The nucleic acid composition of embodiment 38, wherein the antibody-based therapeutic is an antibody, a functional fragment of an antibody, a chimeric antigen receptor (CAR), or a T cell receptor (TCR). 41. The cytokine is a lymphokine, a monokine, a polypeptide hormone, a growth hormone, a parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, a glycoprotein hormone, a follicle-stimulating hormone (FSH), a thyroid-stimulating hormone (TSH), a luteinizing hormone (LH), a hepatocyte growth factor, a fibroblast growth factor, a prolactin, a placental lactogen, a tumor necrosis factor-α and -β, a Mullerian inhibitory substance, a mouse gonadotropin-related peptide, an inhibin, an activin, a vascular endothelial growth factor, an integrin, a thrombopoietin (TPO), a nerve growth factor, a NGF-β , platelet growth factor, transforming growth factor (TGF), TGF-α, TGF-β, insulin-like growth factor-I and -II, erythropoietin (EPO), osteoinductive factor, interferon, e.g., interferon-α, -β, and -γ, colony stimulating factor (CSF), macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF), granulocyte CSF (GCSF), interleukin (IL), tumor necrosis factor, TNF-α, TNF-β, LIF, or Kit ligand (KL), or a combination of two or more thereof. 42. The nucleic acid composition of the preceding embodiment, wherein the exogenous polynucleotide is up to or about 10,000 nucleotides in length. 43. The nucleic acid composition of the preceding embodiment, wherein the components of the nucleic acid composition are expressed on one or more separate nucleic acids. 44. A nucleic acid composition according to any preceding embodiment, further comprising, in operably linked relation, one or more expression control elements which effect expression of the nucleic acid composition in vitro or in vivo. 45. The nucleic acid composition of embodiment 44, wherein the expression control element is a promoter that drives expression of the nucleic acid complex in vitro. 46. ​​The nucleic acid composition of embodiment 45, wherein the promoter is a T7, T3, SP6, or any phage promoter. 47. The nucleic acid composition of embodiment 44, wherein the expression control element is a promoter that drives expression of the nucleic acid complex in a target cell. 48. The nucleic acid composition of embodiment 47, wherein the promoter is a CMV, SV40, or any eukaryotic promoter. 49. The nucleic acid composition of embodiment 47 or embodiment 48, wherein the target cell is a neuronal or non-neuronal cell, VERO, COS, CHO, C6 / 36, HeLa, HEK, or HepG2. 50. The nucleic acid composition of embodiment 47 or embodiment 48, wherein the target cell is an oligodendrocyte, a microglia, or an astrocyte. 51. A pharmaceutical composition comprising the nucleic acid composition according to the previous embodiment, or a pharma- ceutically acceptable salt or derivative thereof. 52. A recombinant Zika virus or Zika virus-like particle produced from expressing a nucleic acid composition of any one of embodiments 1-51 in a producer cell, wherein the producer cell has been further infected with a second Zika virus such that the Zika virus C or a derivative of the Zika virus C, the Zika virus prM / M or a derivative of the Zika virus prM / M, and the Zika virus E or a derivative of the Zika virus E are present in the recombinant Zika virus or Zika virus-like particle. 53. The recombinant Zika virus or Zika virus-like particle of embodiment 52, wherein the second Zika virus is a wild-type Zika virus. 54. The recombinant Zika virus or Zika virus-like particle of embodiment 53, wherein the wild-type Zika virus is an African, Asian, or Brazilian strain. 55. The recombinant Zika virus or Zika virus-like particle of embodiment 53, wherein the second Zika virus is a modified Zika virus. 56. The recombinant Zika virus or Zika virus-like particle described in embodiment 55, wherein the modified Zika virus comprises one or more microRNA-based gene silencing mechanisms. 57. The recombinant Zika virus or Zika virus-like particle of embodiment 56, wherein the one or more microRNA-based gene silencing mechanisms control viral replication. 58. A recombinant Zika virus or Zika virus-like particle produced from expressing a nucleic acid composition of one of embodiments 1-51 in a producer cell that is not infected with a second Zika virus, wherein the Zika virus C or a derivative of the Zika virus C, the Zika virus prM / M or a derivative of the Zika virus prM / M, and the Zika virus E or a derivative of the Zika virus E are present in the recombinant Zika virus or Zika virus-like particle. 59. The recombinant Zika virus or Zika virus-like particle of one of embodiments 52-58, wherein the producer cells are Vero E6, HEK, HEK 293T, HEK 293TT, FreeStyle™ 293-F cells, HEK-293.2sus, or C6 / 36 cells. 60. A recombinant Zika virus or Zika virus-like particle described in one of embodiments 52 to 59, wherein the recombinant Zika virus is replication-competent. 61. A recombinant Zika virus or Zika virus-like particle according to one of embodiments 52 to 60, wherein the recombinant Zika virus is unable to replicate without reducing vector titer or impairing expression of an exogenous polynucleotide. 62. A recombinant Zika virus or Zika virus-like particle described in one of embodiments 52 to 61, wherein the recombinant Zika virus has reduced insertional mutagenesis. 63. A recombinant Zika virus or Zika virus-like particle according to one of embodiments 52 to 62, wherein the recombinant Zika virus reduces the immune response. 64. A pharmaceutical composition comprising a recombinant Zika virus or Zika virus-like particle according to one of embodiments 52-63, and a pharma- ceutical acceptable salt or derivative thereof. 65. A method for delivering an exogenous polynucleotide to a target cell, comprising administering to the target cell a recombinant Zika virus or Zika virus-like particle comprising the exogenous polynucleotide described in one of embodiments 52 to 64. 66. The method of embodiment 65, wherein the target cell is a neuronal cell or a non-neuronal cell. 67. The method of embodiment 66, wherein the neuronal cells are oligodendrocytes, microglia, or astrocytes. 68. The method of embodiment 66, wherein the non-neuronal cell is a prostate epithelial cell, a urethral epithelial cell, a Sertoli cell, a Leydig cell, a spermatogonium cell, or a retinal cell. 69. The method according to one of embodiments 65 to 68, which is carried out in vitro, ex vivo or in vivo. 70. The method of any one of embodiments 65 to 69, wherein the target cells transiently express the exogenous polynucleotide after delivery. 71. The method of any one of embodiments 65 to 69, wherein the target cells persistently express the exogenous polynucleotide after delivery. 72. A method for treating Alzheimer's disease in a subject in need of such treatment, comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described in one of embodiments 52-71, comprising a polynucleotide encoding brain-derived neurotrophic factor (BDNF). 73. A method for treating autism in a subject in need thereof, comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described in any one of embodiments 52-71, comprising a polynucleotide encoding an SH3 and multiple ankyrin repeat domains protein (SHANK). 74. A method for treating Parkinson's disease in a subject in need of such treatment, comprising administering to the subject an effective amount of a recombinant Zika virus or Zika virus-like particle described in one of embodiments 52-71, comprising a polynucleotide encoding aromatic L-amino acid decarboxylase (AADC). 75. The method of one of embodiments 65-74, wherein the administering step is by systemic delivery. 76. The method of one of embodiments 65 to 74, wherein the administering step is performed intravenously and / or intratumorally. 77. The method of any one of embodiments 65 to 76, wherein the administering step targets ventricular, cisternal, subpial, and / or intrathecal cerebrospinal fluid. 78. The method of any one of embodiments 65 to 77, wherein the administering step is not performed intraparenchymal. 79. A nucleic acid composition comprising: (i) a polynucleotide encoding a ribozyme; (ii) a 5' untranslated region (UTR) of a Zika virus; (iii) a 3' UTR of a Zika virus; and (iv) a polynucleotide exogenous to a Zika virus, Optionally, the polynucleotide encoding the ribozyme is located before the 5' UTR or after the 3' UTR; Further optionally, the polynucleotide encoding the ribozyme is a first polynucleotide encoding a ribozyme, and the nucleic acid composition comprises the second polynucleotide, wherein the first polynucleotide is located before the 5' UTR and the second polynucleotide is located after the 3' UTR. 80. The nucleic acid composition of embodiment 79, wherein the ribozyme is a hammerhead ribozyme or a hepatitis D virus (HDV) ribozyme. 81. The nucleic acid composition of embodiment 79 or embodiment 80, further comprising a polynucleotide encoding a second ribozyme. 82. The nucleic acid composition of embodiment 81, wherein the second ribozyme is a hammerhead ribozyme or a hepatitis D virus (HDV) ribozyme. 83. The nucleic acid composition of embodiment 81 or embodiment 82, wherein the polynucleotide encoding the ribozyme encodes a hammerhead ribozyme and the polynucleotide encoding the second ribozyme encodes an HDV ribozyme. 84. The nucleic acid composition of any one of embodiments 79 to 83, comprising a C portion (e.g., a truncation of C, e.g., less than 30, less than 25, less than 20, less than 15, less than 10, or less than 5 amino acids of C, or about 30, about 25, about 20, about 15, about 10, or about 5 amino acids of C). 85. The nucleic acid composition of any one of embodiments 79 to 84, wherein the Zika virus is an African Zika virus, an Asian Zika virus, a component of a Brazilian Zika virus, or a combination of one or two of these. 86. The nucleic acid composition of any one of embodiments 79 to 85, wherein the Zika virus is the African MR766 strain. 87. The nucleic acid composition according to any one of embodiments 79 to 86, wherein the exogenous polynucleotide is an antigen or an antigenic epitope thereof. 88. The nucleic acid composition of embodiment 87, wherein the antigen or antigenic epitope thereof is disease-associated. 89. The nucleic acid composition of embodiment 88, wherein the antigen or antigenic epitope thereof is derived from a pathogen. 90. The nucleic acid composition of embodiment 89, wherein the pathogen is a virus. 91. The nucleic acid composition of embodiment 90, wherein the virus is a human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papillomavirus (HPV-6, HPV-11), measles virus, rabies virus, poliovirus, or yellow fever virus. 92. The nucleic acid composition of embodiment 89, wherein the pathogen is a bacterium. 93. The bacteria may be Acinetobacter baumannii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans. 94. The nucleic acid composition of embodiment 89, wherein the pathogen is a fungus, an amoeba, or a parasite. 95. The fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 brucei, Trypanosoma cruzi, and Taenia multiceps. 96. The nucleic acid composition of any one of embodiments 79 to 95, wherein the exogenous polynucleotide encodes a gene editing tool. 97. The nucleic acid composition of embodiment 96, wherein the gene editing tool is selected from the group consisting of meganuclease-related agents, CRISPR-related agents, TALEN-related agents, and zinc finger-related agents. 98. The nucleic acid composition according to any one of embodiments 79 to 97, wherein the exogenous polynucleotide is a small interfering RNA (siRNA), an antisense RNA, a microRNA (miRNA), a small or short hairpin RNA (shRNA), a guide RNA (gRNA), a clustered regularly interspaced short palindromic repeat RNA (crRNA), a trans-activating clustered regularly interspaced short palindromic repeat RNA (tracrRNA), an immunostimulatory oligonucleotide, an antisense nucleic acid, or a ribozyme. 99. The nucleic acid composition of embodiment 98, wherein the exogenous polynucleotide targets BACE1 or APP. 100. The nucleic acid composition of any one of embodiments 79 to 99, wherein the exogenous polynucleotide encodes a polypeptide associated with a genetic disorder. 101. The nucleic acid composition of embodiment 100, wherein the polypeptide is brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neprilysin inhibitor (NEP), endothelin-converting enzyme (ECE), cathepsin B (CTSB), apolipoprotein E2 (APOE2), SH3 and multiple ankyrin repeat domains protein (SHANK), neurturin (NRTN), glial cell line-derived neurotrophic factor (GDNF), brain dopamine neurotrophic factor (CDNF), vascular endothelial growth factor A (VEGF-A), or aromatic L-amino acid decarboxylase (AADC). 102. The nucleic acid composition of any one of embodiments 79 to 101, wherein the exogenous polynucleotide encodes a therapeutic or diagnostic agent. 103. The nucleic acid composition of embodiment 102, wherein 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. 104. The nucleic acid composition of embodiment 103, wherein the inhibitor or antagonist of the immune checkpoint regulator is an anti-PD1 antibody. 105. The nucleic acid composition of embodiment 103, 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). 106. The cytokine may be a lymphokine, a monokine, a polypeptide hormone, a growth hormone, a parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, a glycoprotein hormone, a follicle-stimulating hormone (FSH), a thyroid-stimulating hormone (TSH), a luteinizing hormone (LH), a hepatocyte growth factor, a fibroblast growth factor, a prolactin, a placental lactogen, a tumor necrosis factor-α and -β, a Mullerian inhibitory substance, a mouse gonadotropin-related peptide, an inhibin, an activin, a vascular endothelial growth factor, an integrin, a thrombopoietin (TPO), a nerve growth factor, or a NGF-β. , platelet growth factor, transforming growth factor (TGF), TGF-α, TGF-β, insulin-like growth factor-I and -II, erythropoietin (EPO), osteoinductive factor, interferon, e.g., interferon-α, -β, and -γ, colony-stimulating factor (CSF), macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF), granulocyte CSF (GCSF), interleukin (IL), tumor necrosis factor, TNF-α, TNF-β, LIF, or Kit ligand (KL), or a combination of two or more thereof. 107. The nucleic acid composition according to one of embodiments 79 to 106, wherein the exogenous polynucleotide is up to or about 10,000 nucleotides in length. 108. A nucleic acid composition according to one of embodiments 79 to 107, further comprising one or more expression control elements operably linked thereto that effect expression of the nucleic acid composition in vitro. 109. The nucleic acid composition of embodiment 108, wherein the expression control element is a promoter that drives expression of the nucleic acid complex in vitro. 110. The nucleic acid composition of embodiment 109, wherein the promoter is a T7, T3, SP6, or any phage promoter. 111. A pharmaceutical composition comprising a nucleic acid composition according to one of embodiments 79 to 110, or a pharma- ceutically acceptable salt or derivative thereof. 112. A method for delivering an exogenous polynucleotide to a target cell, comprising applying to the target cell a compound produced from a nucleic acid composition described in one of embodiments 79 to 111, comprising the exogenous polynucleotide, optionally wherein the compound has a homing property to the target cell, optionally wherein the target cell is a cell of the central nervous system, and optionally wherein the target cell is present in a subject, and wherein applying to the target cell comprises administering the compound to the subject, and optionally wherein the subject is a human or non-human animal. 113. The method of embodiment 112, wherein the compound is produced by encapsulating a transcript produced from the nucleic acid composition using a lipid-based agent. 114. The method of embodiment 113, wherein the transcript is uncapped. 115. The method of embodiment 113 or embodiment 114, wherein the transcript is produced by transcribing the nucleic acid composition in vitro. 116. The method of any one of embodiments 112 to 115, wherein the lipid-based agent is a lipofectamine-related reagent, a liposome, or a lipid nanoparticle. 117. A method for inducing or boosting an immune response in a subject, comprising administering to the subject an effective amount of a nucleic acid composition described in any one of embodiments 79 to 110 or a compound produced from the composition described in embodiment 11, comprising an antigen or an epitope thereof, optionally wherein the subject is a human or a non-human animal. 118. The method of embodiment 117, wherein the compound is produced by encapsulating a transcript produced from the nucleic acid composition using a lipid-based agent. 119. The method of embodiment 118, wherein the transcript is uncapped. 120. The method of embodiment 188 or embodiment 119, wherein the transcript is produced by transcribing the nucleic acid composition in vitro. 121. The method of any one of embodiments 117 to 120, wherein the lipid-based agent is a lipofectamine-related reagent, a liposome, or a lipid nanoparticle. 122. The method of any one of embodiments 112-121, wherein the administering step is performed intramuscularly. 123. The method of any one of embodiments 112-122, wherein the antigen or its antigenic epitope is derived from a pathogen. 124. The method of embodiment 123, wherein the pathogen is a virus. 125. The method of embodiment 124, wherein the virus is a human SARS coronavirus, influenza A virus, influenza B virus, influenza C virus, Ebola virus, hepatitis B virus, hepatitis C virus, herpes simplex virus, human immunodeficiency virus (HIV), human papillomavirus (HPV-6, HPV-11), measles virus, rabies virus, poliovirus, or yellow fever virus. 126. The method of embodiment 123, wherein the pathogen is a bacterium. 127. The bacteria may be Acinetobacter baumannii, Aggregatobacter actinomycetemcomitans, Bartonella bacilliformis, Bartonella henselae, Bartonella quintana, Bifidobacterium, Borrelia, Bortadella pertussis, Brucella sp, Burkholderia cepacis, Burkholderia psedomallei, Campylobacter jejuni, 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 nucleatum, Haemophilus influenza, Haemophilus aphrophilus, Haemophilus ducreyi, Haemophilus parainfluenzaeparainfluenzae, 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 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 cholerae choleraesuis, Serratia marcescens, Schigella dysenteriae, Schigella flexneri, Schigella boydii, Schigella sonneisonnei, 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 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 aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Staphylococcus aureus (VRSA), Staphylococcus lugdunensis, Staphylococcussaprophyticus, Streptococcus pneumonia, Streptococcus pyogenes, or Streptococcus mutans. 128. The method of embodiment 123, wherein the pathogen is a fungus, an amoeba, or a parasite. 129. The fungus, amoeba, or parasite is selected from the group consisting of Acanthamoeba spp., American trypanosomiasis, 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 brucei, Trypanosoma cruzi, and Taenia multiceps.

[0262] [Table 1-1]

[0263]

Table 1-2

[0264]

Table 1-3

[0265]

Table 1-4

[0266]

Table 1-5

[0267]

Table 1-6

[0268]

Table 1-7

[0269]

Table 1-8

[0270]

Table 1-9

[0271]

Table 1-10

[0272]

Table 1-11

[0273]

Table 1-12

[0274]

Table 1-13

[0275]

Table 1-14

[0276]

Table 1-15

[0277]

Table 1-16

[0278]

Table 1-17

[0279]

Table 1-18

[0280]

Table 1-19

[0281]

Table 1-20

[0282]

Table 1-21

[0283]

Table 1-22

[0284]

Table 1-23

[0285]

Table 1-24

[0286]

Table 1-25

[0287]

Table 1-26

[0288]

Table 1-27

[0289]

Table 1-28

[0290]

Table 1-29

[0291]

Table 1-30

[0292]

Table 1-31

[0293]

Table 1-32

[0294]

Table 1-33

[0295]

Table 1-34

[0296]

Table 1-35

[0297]

Table 1-36

[0298]

Table 1-37

[0299]

Table 1-38

[0300]

Table 1-39

[0301]

Table 1-40

[0302]

Table 1-41

[0303]

Table 1-42

[0304]

Table 1-43

[0305]

Table 1-44

[0306]

Table 1-45

[0307]

Table 1-46

[0308]

Table 1-47

[0309]

Table 1-48

[0310]

Table 1-49

[0311]

Table 1-50

[0312]

Table 1-51

[0313]

Table 1-52

[0314]

Table 1-53

[0315]

Table 1-54

[0316]

Table 1-55

[0317]

Table 1-56

[0318]

Table 1-57

[0319]

Table 1-58

[0320]

Table 1-59

[0321]

Table 1-60

[0322]

Table 1-61

[0323]

Table 1-62

[0324]

Table 1-63

[0325]

Table 1-64

[0326]

Table 1-65

[0327]

Table 1-66

[0328]

Table 1-67

[0329]

Table 1-68

[0330]

Table 1-69

[0331]

Table 1-70

[0332]

Table 1-71

[0333]

Table 1-72

[0334]

Table 1-73

[0335]

Table 1-74

[0336]

Table 1-75

[0337]

Table 1-76

[0338]

Table 1-77

[0339]

Table 1-78

[0340]

Table 1-79

[0341]

Table 1-80

[0342]

Table 1-81

[0343]

Table 1-82

[0344]

Table 1-83

[0345]

Table 1-84

[0346]

Table 1-85

[0347]

Table 1-86

[0348]

Table 1-87

[0349]

Table 1-88

[0350]

Table 1-89

[0351]

Table 1-90

[0352]

Table 1-91

[0353]

Table 1-92

[0354]

Table 1-93

[0355]

Table 1-94

[0356]

Table 1-95

[0357]

Table 1-96

[0358]

Table 1-97

[0359]

Table 1-98

[0360]

Table 1-99

[0361]

Table 1-100

[0362]

Table 1-101

[0363]

Table 1-102

[0364]

Table 1-103

[0365]

Table 1-104

[0366]

Table 1-105

[0367]

Table 1-106

[0368]

Table 1-107

[0369]

Table 1-108

[0370]

Table 1-109

[0371]

Table 1-110

[0372]

Table 1-111

[0373]

Table 1-112

[0374] [Table 1-113]

[0375] [Table 1-114]

[0376] [Table 1-115]

[0377] [Table 1-116]

[0378] [Table 1-117]

[0379] [Table 1-118]

[0380] [Table 1-119] EXAMPLES

[0381] Example 1: In vitro transcription and RNA transfection of constructs carrying Nluc and eGFP constructs in Vero cells

[0382] The construct of the plasmid carrying the sequence of RNA polymerase promoter, hammerhead ribozyme, 5'UTR, C part, 3'UTR, HDV ribozyme, and encoding GFP and Nluc luciferase was designed as shown in FIG. 5. See also Table 2.

[0383] RNA generated by in vitro transcription of linearized plasmids was transfected into Vero cells. Transient transfection was performed using Lipofectamine™ MessengerMAX™ (Thermo Scientific™ Catalog No. LMRNA001) and 24 hours later, cells were fed with fresh culture medium.

[0384] After 48 hours, transfected Vero cells were imaged live. GFP signals were detected using a fluorescent microscope (EVOS M5000 Imaging System, ThermoFisher™). As shown in Figure 3, the GFP signal was strong in positively transfected Vero cells after GFP expression by cytoplasmic RNA replicon transfection.

[0385] In a separate experiment, Nluc luciferase activity was determined after transfection with uncapped RNA.

[0386] Plasmids with Nluc activity were transcribed in vitro using the kit MEGAscript (catalog no. AM1330, Life Technologies) according to the user manual. Briefly, 1 μg of linearized plasmid was used in the reaction for 2 h. After precipitation with lithium chloride and isopropanol, the RNA was resuspended in water and stored at -80°C.

[0387] The transcripts were pretreated with Lipofectamine™ MessengerMAX™ (Thermo Scientific™ Catalog No. LMRNA001) according to the manufacturer's recommended conditions. Briefly, the complexes consisting of the transcripts and Lipofectamine™ MessengerMAX™ were incubated with Vero cells (obtained from ATCC), and after 24 hours, fresh culture medium was added to the cells.

[0388] Supernatants and pellets from transfected cells were resuspended in 1 volume of NanoGlo detection reagent (Promega) previously diluted 50-fold in water. Luminescence was quantified for 0.5 seconds in a GloMax luminometer (Promega). As shown in Figure 6D and Figure 6E, high Nluc activity was observed in both cell lysates and supernatants of Vero cells, demonstrating that uncapped RNAs bearing the ZIKV 5'UTR and 3'UTR are recognized by the human translational machinery.

[0389] [Table 2-1]

[0390] [Table 2-2]

[0391] Example 2: Construction of recombinant Zika virus A plasmid construct was designed that harboured the sequences of a eukaryotic promoter, a hammerhead ribozyme, a 5'UTR, a C portion, a 3'UTR, a HDV ribozyme, and encoded GFP and Nluc luciferase.

[0392] Vero cells were obtained from ATCC and grown under high DMEM culture medium. After reaching 80% confluency, DNA linear plasmids were transfected. Transfection was performed using Lipofectamine P3000 (Thermo Scientific™ Catalog No. L3000015). After 24 hours, fresh culture medium was added to the cells. Zeomicyn [1 mg / mL] was added to the medium to select for stable expression of pREP. This resulted in Vero cells expressing the pREP construct (Vero_pREP) (see Figure 4A).

[0393] As shown in Fig. 4B, Vero_pREP was infected with wild-type Zika virus from a Brazilian strain isolated from a clinical case in Brazil, provided by the Evandro Chagas Institute (Belem, Pará, Brazil). Vero_pREP cells were infected with a virus concentration of MOI = 2. After a 30 min incubation period, the culture infection buffer was replaced with fresh medium.

[0394] After 24, 48, and 72 hours, recombinant Zika virus (ZIKVV) containing the viral vector was produced from the transfected Vero_pREP and harvested from the culture medium.

[0395] The culture supernatant containing ZIKVV was stored at -80°C.

[0396] Separately, the medulloblastoma tumor cell line Daoy, obtained from ATCC™, was cultured in 6-well plates under ATCC culture conditions.

[0397] Medulloblastoma tumor cell lines were infected with the viral vectors generated from the transfected Vero_pREP (see FIG. 4C).

[0398] After 24 hours, the infected medulloblastoma tumor cell line (Daoy) was imaged live. GFP signals were detected using a fluorescent microscope (EVOS M5000 Imaging System, ThermoFisher™). As shown in Figure 4D, GFP fluorescence was observed in the medulloblastoma cell line after infection with the recombinant Zika virus.

[0399] Figure 4E shows medulloblastoma cell lines infected with wild-type Brazilian Zika virus 48 hours later, and Figure 4F shows medulloblastoma cell lines infected with recombinant Zika virus 48 hours later with low numbers of attached cells and cell viability, indicating enhanced oncolytic effect of recombinant Zika virus compared to wild-type Zika virus.

[0400] In another experiment, the vector in FIG. 5 was transcribed in vitro and transfected in Vero cells (step 1 in FIG. 7), followed by infection with ZIKV (step 2 in FIG. 7). 120 hours after infection with ZIKV, the culture supernatant containing the viral vector (ZIKVV) produced in Vero cells was collected and concentrated by PEG centrifugation. To test the functionality of the viral vector, the concentrated supernatant was contacted with target cells (Vero and Daoy cell lines) to confirm whether Nanoluc was delivered by ZIKVV (step 3 in FIG. 7). The graph in step 4 in FIG. 7 shows the quantification of Nanoluc in Daoy and Vero cells after contact with the viral vector (pREP+ZIKV) compared to the pREP control (subjected only to pREP transfection) and ZIKV (subjected only to ZIKV infection). pREP+ZIKV shows higher levels of NanoLuc expression compared to the control group. These results demonstrated that ZIKVV delivered NanoLuc to target cells and confirmed reporter gene expression in the target cells.

[0401] Example 3: Expression of Nluc or eGFP after infection with recombinant Zika virus Medulloblastoma tumor cell lines are infected with recombinant Zika virus prepared as in Example 2. Viability and in vitro transduction efficiency are measured. Briefly, medulloblastoma cell lines are cultured in 96-well plates and infected with recombinant Zika virus at MOI=1. CellTiter-Glo® 2.0 Cell Viability Assay (Promega-Cat. No. G9242) and Nano-Glo® Luciferase Assay (Promega-Cat. No. N1110) are used to measure cell viability and Nluc expression, respectively, according to the manufacturer's instructions.

[0402] Separately, the entire genome of the infected cell line will be sequenced to assess the possibility of insertional mutagenesis.

[0403] Similarly, recombinant Zika virus will be injected into wild-type BalBc mice. Over the course of several weeks after injection, weight, activity, inflammation levels near the injection site and systemic inflammatory markers in the blood, and susceptibility will be monitored. In vivo transduction efficiency and blood-brain barrier penetration efficiency will also be assessed.

[0404] Primary T cells are collected from peripheral blood mononuclear cells (PBMCs) of healthy donor patients, transduced with recombinant Zika virus prepared as in Example 1 or 2, and then the survival rate and in vitro transduction efficiency of the primary T cells after infection are measured.

[0405] Example 4: Treatment of Alzheimer's Disease with Recombinant Zika Virus or Zika Virus-Like Particles Establish an Alzheimer's mouse model.

[0406] Recombinant Zika virus or Zika virus-like particles carrying a polynucleotide encoding brain-derived neurotrophic factor (BDNF) are prepared as described in Examples 1 or 2.

[0407] One month after infection with different titers of recombinant Zika virus or Zika virus-like particles, BDNF-treated and mock-treated mice will be tested in the Morris water maze test and other behavioral tests of memory formation and retention.

[0408] Example 5: RNA quantification of generated viral vectors RNA quantification was performed from the different viral vector batches produced.

[0409] RT-PCR was performed using primers specific for the ZIKV genome (gene ZIKV) and pREP (gene pREP) to compare the RNA amounts of ZIKV genome and pREP after pREP transfection and ZIKV infection in Vero cells (Figure 7, steps 1-3). When comparing pREP RNA copies to ZIKV genome RNA copies of one batch generated after infection with ZIKV at MOI = 1, the difference is 10-fold larger (Figure 8). A second batch generated after infection with ZIKV at MOI = 0.1 shows pREP RNA copy numbers comparable to the ZIKV genome copy numbers (Figure 8). These findings demonstrate that at high MOI, ZIKV has 10-fold more pREP gene copies than the ZIKV genome and that the pREP gene is amplified by a viral mechanism.

[0410] Example 5: In vitro tropism of viral vectors The in vitro tropism of ZIKVV (ZIKV genome and pREP RNA) was evaluated. The entry of ZIKVV, ZIKV wild type, or control (pREP mRNA) was tested in cells susceptible to wild type ZIKV (hCMEC and Daoy), but not in cells insensitive to wild type ZIKV (HCT-8). The cell line was combined with wild type virus (ZIKV wild type), viral vector (ZIKVV), and pREP mRNA (control). If the viral vector retains the ability to enter the cell line as wild type ZIKV does, an mRNA with a reporter gene encoding the enzyme NanoLuc luciferase is delivered and translated, which can be detected by a luminescence assay. ZIKVV can deliver genes of interest to cell lines, including the insensitive HCT-8 cell line, indicating a higher spectrum of action of the viral vector that is not limited to neural and stem-like cells that are susceptible to wild type ZIKV (Figure 9).

[0411] Example 6: Evaluation of ZIKV for delivering various genes of interest mRNA prepared from Zika vectors carrying various genes of interest was evaluated. Examples of vectors are shown in Figure 10 (pREP-Luc / Gal) and Figure 11 (pREP-Spike), where the genes in Figure 10 are polycistronic luciferase and β-galactosidase, and the gene in Figure 11 encodes the SARS-CoV-2 spike protein. Vero cells were transfected with pREP-Luc / Gal, and expression of both reporter genes indicates that the pREP construct has the ability to deliver and drive expression of multiple genes up to 6Kb (Figure 12). Furthermore, the SARS-CoV-2 spike coding sequence was inserted into the pREP plasmid and expression intensity was evaluated using ELISA (Figure 13), indicating that the pREP construct can deliver and drive expression of other genes of interest, confirming the use of pREP mRNA as a vaccine product.

[0412] Example 7: Evaluation of Zika Capsid Coding Sequences Five constructs (Figures 14A-14E) of plasmids that retain the sequences of eukaryotic promoter, RNA polymerase promoter, hammerhead ribozyme, 5'UTR, C portion (15, 30, 45, 60, or 75 bp), 3'UTR, HDV ribozyme, and encode GFP and Nluc luciferase were used to evaluate the Zika capsid sequence required to encapsulate pREP (Figure 5). For the sequence of the C portion, see Tables 2 and 3 below.

[0413] The vector was transcribed in vitro using the kit MEGAscript (catalog number AM1330, Life Technologies) according to the user manual.

[0414] Vero cells were obtained from ATCC and grown under high DMEM culture medium. After reaching 80% confluency, they were transfected with RNA (Figures 14A-14E) and DNA (Figures 14A and 14C) plasmids. DNA transfection was performed using Lipofectamine P3000 (Thermo Scientific™ Catalog No. L3000015) and RNA transfection was performed using Lipofectamine™ MessengerMAX™ (Thermo Scientific™ Catalog No. LMRNA001) based on the manufacturer's instructions.

[0415] After 24 hours, Vero cells were infected with wild-type Zika virus. Vero cells were infected with a virus concentration of MOI=0.01. After a 30 minute incubation period, the culture infection buffer was replaced with fresh medium.

[0416] 120 hours after infection with ZIKV, culture supernatants containing the viral vector (ZIKV) produced in Vero cells were harvested and concentrated by PEG centrifugation.

[0417] RNA quantification was performed from the different viral vectors generated (Figures 14A-14E).

[0418] RT-PCT was performed using primers specific for the ZIKV genome (gene ZIKV) and pREP (gene pREP) to detect the RNA levels of ZIKV genome and pREP after pREP transfection and ZIKV infection in Vero cells. pREP RNA copies were quantified from all pREPs with different sizes of capsid sequences (Figures 16A-16B). Figure 16A shows the efficiency of pREP RNA encapsulation in all tested plasmids (C moieties are 15, 30, 45, 60, and 75 bp). The higher efficiency of pREP RNA encapsulation in plasmids with C moieties of 15, 30, 45, and 60 bp indicates that ZIKVV carrying pREP RNA with capsids of 20 or fewer amino acids is efficiently encapsulated by the viral vector. Figure 16B shows that encapsulation derived from DNA transfection was efficient, and the plasmid with C moiety of 45 bp had a higher amount of pREP RNA.

[0419] To test whether viral vectors encapsidate pREP with different capsid sequence sizes (Figures 14A-14E), concentrated supernatants were contacted with Vero cell lines to confirm whether Nanoluc was delivered by ZIKVV (step 3 in Figure 7). The graphs in (Figures 17A and 17B) show the quantification of Nanoluc in Vero cells after contact with viral vectors carrying 5, 10, 15, 20, and 25 amino acids (pREP+ZIKV) derived from both RNA (Figure 17A) and DNA (Figure 17B) transfections, compared to ZIKV (ZIKV infection only). Viral vectors of all capsid sequence sizes demonstrate higher levels of NanoLuc expression compared to the control group. These results demonstrate that ZIKVV can encapsulate pREP RNA at the C portion (15, 30, 45, 60, and 75 bp) and confirm that NanoLuc is delivered by reporter gene expression in target cells.

[0420] [Table 3]

[0421] Example 8: In vivo biodistribution and tropism of ZIKVV A feature of this technology is the possibility of generating viral vectors with tropism for the CNS and eye. To track the in vivo biodistribution and tropism of ZIKVV, a bioluminescence imaging system was used based on the Nanoluc report gene inserted into the vector gene of interest. ZIKVV was administered systemically to 3-week-old Balb / C male mice by intraperitoneal injection of ZIKVV carrying pREP RNA, generated as in Example 2 (Figure 5). 50 μl of 10-fold diluted ZIKVV was administered with a total of 10^4 particles. For bioluminescence imaging, mice were administered Nanoluc substrate (Promega-50-fold diluted) in a volume of 50 μl by intraperitoneal administration immediately before acquiring images on an IVIS-Spectrum instrument (Perkin-Elmer). Strong Nanoluc signals were observed in the brain and eye regions 48 hours after ZIKVV injection into mice. These results confirm the in vivo tropism of ZIKVV for the eye and brain, and that the vector selectively delivers and transduces the gene of interest to these organs following systemic vector administration.

[0422] The foregoing merely illustrates the principles of the present disclosure. It will be understood that those skilled in the art can devise various arrangements that embody the principles of the present invention and are within its spirit and scope, although not expressly described or shown herein. Furthermore, all examples and conditions described herein are intended primarily to aid the reader in understanding the principles of the present disclosure and the concepts contributed by the inventors to further develop the art, and are not intended to be limited to such specifically described examples and conditions. Furthermore, all descriptions herein of the principles, aspects, and embodiments of the present invention, as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Furthermore, such equivalents are intended to include both currently known equivalents and equivalents developed in the future, i.e., any elements that perform the same function regardless of structure, are developed. Thus, the scope of the present disclosure 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

1. 1. A method for delivering a polynucleotide to a target cell of a subject, the method comprising administering to the subject a Zika virus or Zika virus-like particle comprising the polynucleotide, wherein the polynucleotide is not a Zika virus polynucleotide and the target cell is a cell of the central nervous system.

2. 1. A method for delivering a polynucleotide to a subject, the method comprising administering to the subject a Zika virus or Zika virus-like particle comprising the polynucleotide, wherein the polynucleotide is not a Zika virus polynucleotide and the Zika virus or Zika virus-like particle does not comprise a replicating Zika virus.

3. 1. A method for delivering a polynucleotide to a subject, the method comprising administering to the subject a Zika virus or Zika virus-like particle comprising the polynucleotide, wherein the polynucleotide is not a Zika virus polynucleotide, and the polynucleotide has a length of up to about 10,000 nucleotides.

4. The method of any one of claims 1 to 3, wherein the subject has a disease or disorder of the central nervous system.

5. The method of any one of claims 1 to 3, wherein the polynucleotide has a length of more than about 6000 nucleotides.

6. The method of any one of claims 1 to 3, wherein the subject is a human or non-human animal.

7. 1. A Zika virus or Zika virus-like particle comprising a polynucleotide exogenous to Zika virus having a length of from about 6,000 nucleotides to about 10,000 nucleotides, optionally wherein the polynucleotide does not encode a full-length viral structural protein or a full-length viral nonstructural protein.

8. 8. The Zika virus or Zika virus-like particle of claim 7, wherein the Zika virus or Zika virus-like particle is produced from a nucleic acid comprising the polynucleotide.

9. 9. The Zika virus or Zika virus-like particle of claim 8, wherein the nucleic acid further comprises a Zika virus 5'UTR and a Zika virus 3'UTR.

10. 10. The Zika virus or Zika virus-like particle of claim 9, wherein the Zika virus 5'UTR is at least 90% homologous to or identical to a Zika virus 5'UTR of Table 2, Table 1A, Table 1C, or Table 1H.

11. 10. The Zika virus or Zika virus-like particle of claim 9, wherein the Zika virus 3'UTR is at least 90% homologous to or identical to a Zika virus 3'UTR of Table 2, Table 1A, Table 1B, or Table 1G.

12. 9. The Zika virus or Zika virus-like particle of claim 8, wherein the nucleic acid further comprises a polynucleotide encoding a first ribozyme.

13. 13. The Zika virus or Zika virus-like particle of claim 12, wherein the nucleic acid further comprises a polynucleotide encoding a second ribozyme.

14. 14. The Zika virus or Zika virus-like particle of any one of claims 8-13, wherein the nucleic acid comprises a polynucleotide encoding a derivative of Zika virus C, wherein the derivative of Zika virus C is a truncation of about 5, 10, 15, 20, 25, or 30, or less than about 5, 10, 15, 20, 25, or 30 amino acids of Zika virus C.

15. 14. The Zika virus or Zika virus-like particle of any one of claims 8 to 13, wherein the nucleic acid does not comprise a polynucleotide encoding the Zika virus capsid protein (C) or a derivative of the Zika virus C.

16. 14. The Zika virus or Zika virus-like particle of any one of claims 8 to 13, wherein the nucleic acid does not include a polynucleotide encoding a Zika virus membrane protein (prM / M) or a derivative of the Zika virus prM / M.

17. 14. The Zika virus or Zika virus-like particle of any one of claims 8 to 13, wherein the nucleic acid does not include a polynucleotide encoding the Zika virus envelope protein (E) or a derivative of the Zika virus E.

18. 14. The Zika virus or Zika virus-like particle of any one of claims 8-13, wherein the nucleic acid does not include a polynucleotide encoding one or more nonstructural (NS) proteins: (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) two or more of (i)-(vii).

19. 14. The Zika virus or Zika virus-like particle of any one of claims 8 to 13, wherein the nucleic acid further comprises a promoter.

20. 14. The Zika virus or Zika virus-like particle of any one of claims 8-13, wherein the nucleic acid comprises a sequence of Table 1A-1K, Table 2, or Table 3.

21. 1. A nucleic acid composition comprising: (i) a 5' untranslated region (5'UTR) of a first Zika virus; (ii) a 3'UTR of a second Zika virus, wherein the first Zika virus is optionally identical to the second Zika virus; and (iii) a polynucleotide exogenous to the first Zika virus and the second Zika virus.

22. 22. The nucleic acid composition of claim 21, wherein the polynucleotide is from about 5,000 bases to about 10,000 bases in length.

23. The nucleic acid composition of claim 21 or claim 22, wherein the nucleic acid does not contain a polynucleotide encoding a Zika virus envelope protein (E) or a derivative of the Zika virus E, and wherein the nucleic acid does not contain a polynucleotide encoding one or more nonstructural (NS) proteins, namely, (i) NS1, (ii) NS2A, (iii) NS2B, (iv) NS3, (v) NS4A, (vi) NS4B, (vii) NS5, or (viii) two or more of (i) to (vii).

24. 23. The nucleic acid composition of claim 21 or claim 22, comprising a sequence of Table 1A-1K, Table 2, or Table 3.