Use of thermal bioswitches in AAV-based gene therapy

Abstract

Disclosed herein include methods, compositions, and kits suitable for use in AAV-based gene therapy. In some embodiments, the method comprises administering a composition comprising AAV vectors comprising thermally actuatable genetic circuits encoding therapeutic gene products. In some embodiments, the methods and compositions disclosed herein can be used for treating a retinal disease or disorder in a subject.

Description

USE OF THERMAL BIOSWITCHES IN AAV-BASED GENE THERAPY CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of, and priority to, U.S. Provisional Application 63 / 610,545, filed on December 15, 2023, the contents of which are hereby incorporated herein by reference in their entirety.INCORPORATION BY REFERENCE OF SEQUENCE LISTING

[0002] The contents of the electronic sequence listing (238872000240SEQLIST.xml; Size: 40,250 bytes; and Date of Creation: December 11, 2024) is herein incorporated by reference in its entirety.BACKGROUNDField

[0003] The present disclosure relates generally to the field of gene therapy.Description of the Related Art

[0004] AAV-based gene therapy is becoming more established, with recent approvals for AAV-based therapies including Luxturna for retinitis pigmentosa, Hemgenix for Hemophilia B, and Zolgensma for spinal muscular atrophy. Many clinical trials are ongoing for additional indications. In addition to gene replacement, drug factory applications are being explored. However, AAV therapies can typically only be dosed once (due to anti-vector immunity), making it challenging to subsequently boost dose of therapy. At the same time, too high a dose may result in toxicity. For systemic therapy, tissue tropism (e.g. liver transduction) limits the dose that can be delivered to target tissues. On-going concerns of AAV-based therapies include lack of tunability and forever-on production of therapeutic agents. There is a need for inducible methods for activating or boosting the production of AAV-encoded biologies to enable better control of dosing in a single tissue and / or reduce off-target drug production.SUMMARY

[0005] Disclosed herein include compositions for treating a retinal disease or disorder in a subject. In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product; wherein the first inducible promoter is capable of inducing transcription of the gene to generate the therapeutic gene product upon thermal stimulation.

[0006] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a transactivator gene, and a second promoter operably linked to a second polynucleotidecomprising a therapeutic gene, wherein the first inducible promoter is capable of inducing transcription of the transactivator gene to generate a transactivator transcript in the presence of thermal stimulation, wherein the transactivator transcript is capable of being translated to generate a transactivator; and wherein, in the presence of the transactivator and; optionally, a transactivator-binding compound, the second promoter is capable of inducing transcription of a therapeutic gene to generate a therapeutic gene product.

[0007] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter and a second promoter each operably linked to a first polynucleotide comprising a therapeutic gene and to a second polynucleotide comprising a transactivator gene, wherein the first inducible promoter is capable of inducing transcription of the therapeutic gene and the transactivator gene to generate a polycistronic transcript upon thermal stimulation, wherein, in the presence of the transactivator and a transactivator-binding compound, the second promoter is capable of inducing transcription of the therapeutic gene and the transactivator gene to generate a polycistronic transcript, and wherein the polycistronic transcript is capable of being translated to generate a transactivator and a therapeutic protein.

[0008] In some embodiments, the second promoter comprises one or more copies of a transactivator recognition sequence the transactivator is capable of binding to induce transcription, and wherein the transactivator is incapable of binding the transactivator recognition sequence in the absence of the transactivator-binding compound. In some embodiments, the one or more copies of a transactivator recognition sequence comprise one or more copies of a tet operator (TetO). In some embodiments, the transactivator comprises reverse tetracycline-controlled transactivator (rtTA). In some embodiments, the transactivator comprises tetracycline-controlled transactivator (tTA). In some embodiments, the transactivatorbinding compound comprises tetracycline, doxycycline or a derivative thereof. In some embodiments, the first polynucleotide and the second polynucleotide are operably linked to a tandem gene expression element. In some embodiments, the tandem gene expression element is an internal ribosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof.

[0009] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a recombinase gene, wherein the first inducible promoter is capable of inducing transcription of the recombinase gene to generate a recombinase transcript upon thermal stimulation, and wherein the recombinase transcript is capable of being translated to generate a recombinase; asecond promoter and a second polynucleotide comprising a therapeutic gene, wherein, in the absence of a recombination event, the second promoter and the second polynucleotide are not operably linked, wherein the recombinase is capable of catalyzing the recombination event, and wherein the second promoter and the second polynucleotide are operably linked after the recombination event such that the second promoter is capable of inducing transcription of the therapeutic gene to generate a therapeutic gene product.

[0010] In some embodiments, the recombination event comprises removal of a sequence flanked by recombinase target sites or an inversion of a sequence flanked by recombinase target sites. In some embodiments, the second polynucleotide is flanked by recombinase target sites. In some embodiments, prior to the recombination event, the sequence of the therapeutic gene is inverted relative to the promoter.

[0011] The composition can comprise at least one stop cassette situated between the second promoter and the therapeutic gene, wherein the stop cassette comprises one or more stop sequences, and wherein the one or more stop cassettes are flanked by recombinase target sites. In some embodiments, the therapeutic gene product is an RNA transcript and / or protein. In some embodiments, the transcript is an mRNA capable of being translated to generate the protein. In some embodiments, the at least one stop cassette is configured to prevent transcription of the therapeutic gene and / or translation of the therapeutic transcript. In some embodiments, the one or more stop sequences comprise a polyadenylation signal, a stop codon, a frame-shifting mutation, or any combination thereof. In some embodiments, the second promoter comprises a ubiquitous or tissue-specific promoter.

[0012] In some embodiments, the second promoter comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focus-forming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl 1 promoters from vaccinia virus, an elongation factor 1 -alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P-kinesin (P- KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P- actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CBH promoter, any variant thereof, or any combination thereof. In some embodiments, the recombinase is Cre, Dre, Flp, KD, B2, B3, , HK022, HP1, y6, ParA, Tn3, Gin, C31, Bxbl, R4, derivatives thereof, orany combination thereof. In some embodiments, the recombinase is a Flp recombinase and the recombinase target sites are FRT sites or the recombinase is a Cre recombinase and the recombinase target sites are loxP sites.

[0013] In some embodiments, the first inducible promoter comprises a core promoter and at least one heat shock element (HSE). In some embodiments, the core promoter comprises the heat shock protein transcription start site of a gene selected from the group consisting of HSPA1 A, HSPH1, HSPB1, HSPA6, and YB. In some embodiments, the core promoter comprises the sequence of any one of SEQ ID NOs: 9-12 or a sequence having at least 85% identity to any one of SEQ ID NOs: 9-12. In some embodiments, the core promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 9-12. In some embodiments, the first inducible promoter comprises seven HSEs. In some embodiments, each of the at least one HSE comprises the sequence of 5’ - nGAAnnTTCnnGAAn-3’. In some embodiments, the at least one HSE comprises the sequence of any one of SEQ ID NOs: 1-8 or a sequence having at least 85% identity to any one of SEQ ID NOs: 1-8 or a sequence differing by one or two nucleotide mismatches relative to any one of SEQ ID NOs: 1-8. In some embodiments, the at least one HSE comprises a sequence selected from the group consisting of SEQ ID NOs: 1-8. In some embodiments, the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 13-20 or a sequence having at least 85% identity to any one of SEQ ID NOs: 13-20. In some embodiments, the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 13-20. In some embodiments, the first inducible promoter comprises the sequence of any one of SEQ ID NOS: 21-34 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-34. In some embodiments, the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-34.

[0014] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a leaky inducible promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product, wherein the leaky inducible promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the leaky inducible promoter is capable of boosting transcription of the gene, thereby driving a second level of expression of the therapeutic gene product upon thermal stimulation.

[0015] In some embodiments, the leaky inducible promoter comprises an HSPB, HSPB’2, HSP A / A, or HSP A / B promoter. In some embodiments, the leaky inducible promoter comprises the sequence of any one of SEQ ID NOs: 21-24 and 28-29 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-24 and 28-29. In some embodiments, the leaky induciblepromoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-24 and 28-29.

[0016] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a tandem promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product, wherein the tandem promoter comprises a first inducible promoter and a second constitutive promoter, wherein the second constitutive promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the first inducible promoter is capable of boosting transcription of the gene upon thermal stimulation, thereby driving a second level of expression of the therapeutic gene product. In some embodiments, the tandem promoter comprises, from 5’ to 3’: the first inducible promoter then the second constitutive promoter. In some embodiments, the tandem promoter comprises, from 5’ to 3’: the second constitutive promoter then the first inducible promoter.

[0017] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a gene encoding a therapeutic gene product and a second constitutive promoter operably linked to a second polynucleotide comprising the gene encoding a therapeutic gene product wherein the second constitutive promoter induces transcription of the gene encoding a therapeutic gene product from the second polynucleotide in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the first inducible promoter is capable of inducing the transcription of the therapeutic gene from the first polynucleotide upon thermal stimulation, thereby boosting the expression of the therapeutic gene product to a second level of expression.

[0018] In some embodiments, the nucleic acid comprises, from 5’ to 3’ : the first inducible promoter, the first polynucleotide, the second constitutive promoter, and the second polynucleotide. In some embodiments, the nucleic acid comprises, from 5’ to 3’: the second constitutive promoter, the second polynucleotide, the first inducible promoter, and the first polynucleotide. In some embodiments, the second level of expression is at least 1.1-fold higher than the first level of expression. In some embodiments, after a first duration of time, the therapeutic gene product returns to the first level of expression from the second level of expression, wherein the first duration of time is about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks or more.

[0019] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a bidirectional promoter comprising a first inducible promoter in a firstorientation, and a second constitutive promoter in a second orientation, wherein the first inducible promoter is operably linked to a first polynucleotide comprising a first gene encoding a therapeutic gene product, and wherein the second constitutive promoter is operably linked to a second polynucleotide comprising a second gene, wherein the first and second orientations are opposite of each other; wherein the second constitutive promoter is capable of inducing transcription of the second gene in the absence of thermal stimulation; and wherein the first inducible promoter is capable of inducing transcription of the first gene upon thermal stimulation to generate the therapeutic gene product.

[0020] In some embodiments, the second gene encodes a therapeutic gene product. In some embodiments, the therapeutic gene product is capable of inhibiting vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6). In some embodiments, the therapeutic gene product is selected from the group consisting of RPE65, Complement Factor 1 (CF1), CD59, FMS Related Receptor Tyrosine Kinase 1 (FLT-1), Rab escort protein 1 (REP1), NADH hydrogenase 4 (ND4), MER proto-oncogene tyrosine kinase (MERTK), X-linked retinitis pigmentosa GTPase regulator (RPGR), or Retinoschisin (RSI). In some embodiments, the second constitutive promoter comprises a ubiquitous or tissue-specific promoter. In some embodiments, the second constitutive promoter comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focus-forming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl 1 promoters from vaccinia virus, an elongation factor 1 -alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P-kinesin (P- KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P- actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CBH promoter, or any combination thereof.

[0021] In some embodiments, the first inducible promoter comprises a core promoter and at least one heat shock element (HSE). In some embodiments, the core promoter comprises the heat shock protein transcription start site of a gene selected from the group consisting of HSPA1 A, HSPH1, HSPB1, HSPA6, and YB. In some embodiments, the core promoter comprises the sequence of any one of SEQ ID NOs: 9-12 or a sequence having at least 85% identity to any one of SEQ ID NOs: 9-12. In some embodiments, the core promoter comprises a sequence selectedfrom the group consisting of SEQ ID NOs: 9-12. In some embodiments, the first inducible promoter comprises seven HSEs. In some embodiments, each of the at least one HSE comprises the sequence of 5’ - nGAAnnTTCnnGAAn-3’. In some embodiments, the at least one HSE comprises the sequence of any one of SEQ ID NOs: 1-8 or a sequence having at least 85% identity to any one of SEQ ID NOs: 1-8 or a sequence differing by one or two nucleotide mismatches relative to any one of SEQ ID NOs: 1-8. In some embodiments, the at least one HSE comprises a sequence selected from the group consisting of SEQ ID NOs: 1-8. In some embodiments, the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 13-20 or a sequence having at least 85% identity to any one of SEQ ID NOs: 13-20. In some embodiments, the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 13-20. In some embodiments, the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 21-34 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-34. In some embodiments, the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-34. In some embodiments, any two of the first inducible promoter, the second promoter, the leaky inducible promoter, the tandem promoter, the second constitutive promoter, and the bidirectional promoter are in opposite orientations or the same orientation relative to each other.

[0022] In some embodiments, the therapeutic gene product comprises an siRNA, an shRNA, a miRNA, or a protein. In some embodiments, the therapeutic gene product is capable of inhibiting vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6). In some embodiments, the therapeutic gene product comprises an antibody or fragment thereof capable of binding to vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6). In some embodiments, the therapeutic gene product comprises RPE65, Complement Factor 1 (CF1), CD59, FMS Related Receptor Tyrosine Kinase 1 (FLT-1), Rab escort protein 1 (REP1), NADH hydrogenase 4 (ND4), MER proto-oncogene tyrosine kinase (MERTK), X-linked retinitis pigmentosa GTPase regulator (RPGR), or Retinoschisin (RSI).

[0023] In some embodiments, the thermal stimulation comprises heating to an activating temperature, wherein the activating temperature is greater than 37°C. In some embodiments, the activating temperature comprises a temperature of at least 37°C to at most 70°C. In some embodiments, the activating temperature is about 37.0°C, 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, about 46.0°C, about 46.5°C, about 47.0°C, about 47.5°C, about 48.0°C, about 48.5°C, about 49.0°C, about 49.5°C, about 50.0°C, about 50.5°C, about 51.0°C, about 51.5°C, about 52.0°C, about 52.5°C, about 53.0°C, about 53.5°C, about 54.0°C, about 54.5°C, about 55.0°C, about 55.5°C, about 56.0°C, about 56.5°C, about 57.0°C, about 57.5°C, about 58.0°C, about 58.5°C, about 59.0°C, about 59.5°C, about 60.0°C, about 60.5°C, about 61.0°C, about 61.5°C, about 62.0°C, about 62.5°C, about 63.0°C, about 63.5°C, about 64.0°C, about 64.5°C, about 65.0°C, about 65.5°C, about 66.0°C, about 66.5°C, about 67.0°C, about 67.3°C, about 67.5°C, about 68.0°C, about 68.5°C, about 69.0°C, about 69.5°C, or about 70.0°C. In some embodiments, the activating temperature is about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, or about 46.0°C. In some embodiments, the activating temperature comprises a temperature of about 42°C to about 43°C. In some embodiments, the activating temperature is about 42.0°C, about 42.1°C, about 42.2°C, about 42.3°C, about 42.4°C, about 42.5°C, about 42.6°C, about 42.7°C, about 42.8°C, about 42.9°C, or about 43.0°C.

[0024] In some embodiments, the viral vector is or comprises an AAV vector, a lentivirus vector, a retrovirus vector, an adenovirus vector, a herpesvirus vector, a herpes simplex virus vector, a cytomegalovirus vector, a vaccinia virus vector, a MVA vector, a baculovirus vector, a vesicular stomatitis virus vector, a human papillomavirus vector, an avipox virus vector, a Sindbis virus vector, a VEE vector, a Measles virus vector, an influenza virus vector, a hepatitis B virus vector, an integration-deficient lentivirus (IDLV) vector derivatives thereof, or any combination thereof. In some embodiments, the viral vector is capable of integrating into a mammalian cell genome, and wherein the therapeutic gene product is capable of being expressed upon integration. In some embodiments, the viral vector is a lentiviral vector. In some embodiments, the viral vector is human immunodeficiency virus 1 (HIV-1), human immunodeficiency virus 2 (HIV-2), visna-maedi virus (VMV) virus, caprine arthritisencephalitis virus (CAEV), equine infectious anemia virus (EIAV), feline immunodeficiency virus (FIV), bovine immune deficiency virus (BIV), simian immunodeficiency virus (SIV), derivatives thereof, or any combination thereof. In some embodiments, the viral vector is a recombinant lentiviral vector. In some embodiments, the recombinant lentiviral vector is derived from a lentivirus pseudotyped with vesicular stomatitis virus (VSV), lymphocytic choriomeningitis virus (LCMV), Ross river virus (RRV), Ebola virus, Marburg virus, Mokala virus, Rabies virus, RD114, or variants thereof. The composition can comprise one or more of a left (5') retroviral LTR, a Psi ( ) packaging signal, a central polypurine tract / DNA flap (cPPT / FLAP), a retroviral export element, and a right (3') retroviral LTR. In some embodiments,the 5' LTR or 3' LTR is a lentivirus LTR. In some embodiments, the 3' LTR comprises one or more modifications and / or deletions. In some embodiments, the 3' LTR is a self-inactivating (SIN) LTR.

[0025] In some embodiments, the viral vector comprises an AAV vector. In some embodiments, the AAV vector comprises a single-stranded AAV (ssAAV) vector or a self- complementary AAV (scAAV) vector. In some embodiments, the AAV vector comprises AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, derivatives thereof, or any combination thereof. In some embodiments, the AAV vector comprises an AAV9 variant engineered for systemic delivery. In some embodiments, the AAV vector comprises AAVPHP.B, AAV-PHP.eB, or AAV-PHP.S.

[0026] In some embodiments, the AAV vector is or comprises an AAV selected from the group consisting of AAV9, AAV9 K449R (or K449R AAV9), AAV1, AAVrhlO, AAV-DJ, AAV-DJ8, AAV5, AAVPHP.B (PHP.B), AAVPHP.A (PHP. A), AAVG2B-26, AAVG2B-13, AAVTH1.1-32, AAVTH1.1-35, AAVPHP.B2 (PHP.B2), AAVPHP.B 3 (PHP.B3), AAVPHP.N / PHP.B-DGT, AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP, AAVPHP.B-ATT-T, AAVPHP.B-DGT-T, AAVPHP.B-GGT-T, AAVPHP.B-SGS, AAVPHP.B-AQP, AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B-SNP, AAVPHP.B- QGT, AAVPHP.B-NQT, AAVPHP.B- EGS, AAVPHP.B-SGN, AAVPHP.B-EGT, AAVPHP.B- DST, AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP, AAVPHP.B-SQP, AAVPHP.B- QLP, AAVPHP.B- TMP, AAVPHP.B-TTP, AAVPHP.S / G2A12, AAVG2 Al 5 / G2A3 (G2A3), AAVG2B4 (G2B4), AAVG2B5 (G2B5), PHP.S, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11, AAV 12, AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-lb, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa, AAV43-1, AAV43-12, AAV43-20, AAV43-21, AAV43-23, AAV43-25, AAV43-5, AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7, AAV1- 7 / rh.48, AAVl-8 / rh.49, AAV2-15 / rh.62, AAV2-3 / rh.61, AAV2-4 / rh.5O, AAV2-5 / rh.51, AAV3. l / hu.6, AAV3. l / hu.9, AAV3-9 / rh.52, AAV3-1 l / rh.53, AAV4-8 / rl 1.64, AAV4-9 / rh.54, AAV4- 19 / rh.55, AAV5-3 / rh.57, AAV5-22 / rh.58, AAV7.3 / hu.7, AAV16.8 / hu.lO, AAV16.12 / hu.ll, AAV29.3 / bb.l, AAV29.5 / bb.2, AAV106. l / hu.37, AAV1 14.3 / hu.4O, AAV127.2 / hu.41, AAV127.5 / hu.42, AAV128.3 / hu.44, AAV130.4 / hu.48, AAV145. l / hu.53, AAV145.5 / hu.54, AAV145.6 / hu.55, AAV161. 10 / hu.60, AAV161.6 / hu.61, AAV33. 12 / hu.l7, AAV33.4 / hu.l5,AAV33.8 / hu.l6, AAV52 / hu.l9, AAV52.1 / hu.2O, AAV58.2 / hu.25, AAVA3.3, AAVA3.4, AAV A3.5, AAV A3.7, AAVC1, AAVC2, AAVC5, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70, AAVpi.l, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu. 12, AAVH6, AAVH-l / hu.l, AAVH-5 / hu.3, AAVLG-10 / rh.40, AAVLG-4 / rh.38, AAVLG-9 / hu.39, AAVN721- 8 / rh.43, AAVCh.5, AAVCh.5Rl, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5Rl, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.l, AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.10, AAVhu.ll, AAVhu.13, AAVhu.15, AAVhu.16, AAVhu. 1 7, AAVhu.18, AAVhu.20, AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu.31,AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44Rl, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48Rl, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.l 4 / 9, AAVhu.t 19, AAVrh.2, AAVrh.2R, AAVrh.8, AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13, AAVrh.BR, AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49, AAVrh.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64Rl, AAVrh.64R2, AAVrh.67, AAVrh.73, AAVrh.74, AAVrh8R, AAVrh8R A586R mutant, AAVrh8RR533 A mutant, AAAV, BAAV, caprine AAV, bovine AAV, AAVhEl.l, AAVhErl.5, AAVhER1.14, AAVhErl.8, AAVhErl. 16, AAVhErl.18, AAVhErl.35, AAVhErl.7, AAVhErl.36, AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.3 1, AAVhEr2.36, AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01, AAV-LK02, AAV- LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV-LK09, AAV- LK10, AAV-LK11, AAV-LK12, AAV-LK13, AAV-LK14, AAV-LK15, AAV-LK16, AAV-LK17, AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV- PAEC11, AAV-PAEC12, AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV Shuffle 100-1, AAV Shuffle 100-3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM 10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrh.48, AAVhu.19, AAVhu.ll, AAVhu.53, AAV4-8 / rh.64, AAVLG-9 / hu.39, AAV54.5 / hu.23, AAV54.2 / hu.22, AAV54.7 / hu.24, AAV54.1 / hu.21, AAV54.4R / hu.27, AAV46.2 / hu.28, AAV46.6 / hu.29, AAV128. l / hu.43, true type AAV (ttAAV), EGRENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7.10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-El, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV CHt-6. 1, AAV CHt-6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-Pl, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-Bl, AAV CKd-B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV CKd-B8, AAV CKd-Hl, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAV CKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-Fl, AAV CLg-F2, AAV CLg-F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1, AAV CLvl-1, AAV Clvl-10, AAV CLvl-2, AAV CLv-12, AAV CLvl-3, AAV CLv-1 3, AAV CLvl-4, AAV Clvl-7, AAV Clvl-8, AAV Clvl-9, AAV CLv-2, AAV CLv- 3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-Dl, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-El, AAV CLv-Kl, AAV CLv-K3, AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-Ml, AAV CLv-Ml 1, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-Rl, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV CSp-1, AAV CSp- 10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8. 10, AAV CSp-8.2, AAV CSp-8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1, AAVF11 / HSC11, AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5, AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, AAVF9 / HSC9, variants thereof, a hybrid or chimera of any of the foregoing AAV serotypes, or any combination thereof.

[0027] In some embodiments, the AAV vector comprises an AAV variant engineered for reduced binding to heparin sulfate proteoglycans. In some embodiments, the AAV vector comprises an AAV variant engineered for enhanced transduction across the internal limiting membrane (ILM) of the retina. In some embodiments, the AAV vector comprises an AAV variant engineered for enhanced transduction of the retinal cells. In some embodiments, theretinal cells comprise amacrine cells, bipolar cells, rods, cones, cells of the retinal pigment epithelium, or any combination thereof. In some embodiments, the AAV vector comprises a capsid protein comprising an insertion of a targeting peptide after amino acid 587 (AA587) of the AAV capsid protein or a functional equivalent of AA587. In some embodiments, the targeting peptide insertion results in enhanced transduction of retinal cells as compared to an AAV comprising a capsid protein that does not comprise the insertion. In some embodiments, the targeting peptide is about seven amino acids in length. The composition can comprise a recombinant AAV (rAAV). The composition can comprise one or more pharmaceutically acceptable excipients.

[0028] Provided herein are compositions for use in treating a retinal disease or disorder.

[0029] Disclosed herein include methods of treating a retinal disease or disorder in a subject. In some embodiments, the method comprises: administering any of the compositions disclosed herein to the subject; and applying thermal energy to a target site of the subject sufficient to increase local temperature of the target site to an activating temperature, thereby inducing the expression of the therapeutic gene product.

[0030] In some embodiments, the target site comprises the retina. In some embodiments, the composition is administered at a dose of about 1 x 109viral genomes to about 1.0 * 1012viral genomes. In some embodiments, the composition is administered at a dose of more than about 1.0 x 1012viral genomes. In some embodiments, the composition is administered by intravitreal route, subretinal route, or suprachoroidal route.

[0031] In some embodiments, the method comprises a single administration of the composition to the subject. In some embodiments, the method comprises administering the composition two or more times to the subject. In some embodiments, each of the two or more administrations are 1 month, 2 months, 6 months, 1 year, or more, apart.

[0032] In some embodiments, the period of time between the administering the composition and applying thermal energy is about 1 week, about 6 days, about 5 days, about 4 days, about 3 days, about 48 hours, about 44 hours, about 40 hours, about 35 hours, about 30 hours, about 25 hours, 20 hours, 15 hours, 10 hours, about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, about 10 minutes, or about 5 minutes. In some embodiments, the method comprises two or more applications of thermal energy. In some embodiments, the period of time between administering the composition and applying thermal energy is the period of time between the administering the composition and the first application of thermal energy.

[0033] In some embodiments, the activating temperature is greater than 37°C. In some embodiments, the activating temperature comprises a temperature of at least 37.5° C to at most 70°C. In some embodiments, the activating temperature is about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, about 46.0°C, about 46.5°C, about 47.0°C, about 47.5°C, about 48.0°C, about 48.5 °C, about 49.0°C, about 49.5°C, about 50.0°C, about 50.5°C, about 51.0°C, about 51.5°C, about 52.0°C, about 52.5°C, about 53.0°C, about 53.5°C, about 54.0°C, about 54.5°C, about 55.0°C, about 55.5°C, about 56.0°C, about 56.5°C, about 57.0°C, about 57.5°C, about 58.0°C, about 58.5°C, about 59.0°C, about 59.5°C, about 60.0°C, about 60.5°C, about 61.0°C, about 61.5°C, about 62.0°C, about 62.5°C, about 63.0°C, about 63.5°C, about 64.0°C, about 64.5°C, about 65.0°C, about 65.5°C, about 66.0°C, about 66.5°C, about 67.0°C, about 67.3°C, about 67.5°C, about 68.0°C, about 68.5°C, about 69.0°C, about 69.5°C, or about 70.0°C. In some embodiments, the thermal energy is applied to the target site for a duration of time comprising about 1 second, about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, or about 1 minute.

[0034] In some embodiments, the activating temperature is about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, or about 46.0°C. In some embodiments, the thermal energy is applied to the target site for a duration of time comprising about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes.

[0035] In some embodiments, the activating temperature comprises a temperature of about 42°C to about 43°C. In some embodiments, the activating temperature is about 42.0°C, about 42.1°C, about 42.2°C, about 42.3°C, about 42.4°C, about 42.5°C, about 42.6°C, about 42.7°C, about 42.8°C, about 42.9°C, or about 43.0°C.

[0036] In some embodiments, the thermal energy is applied to the target site for a duration of time comprising about 1 second to about 20 minutes.

[0037] In some embodiments, the thermal energy is applied to the target site for a duration of time comprising about 1 second, about 5 seconds, about 10 seconds, about 15 seconds, about 20seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes. In some embodiments, the temperature at the target site is elevated to about 42°C to about 43 °C after the applying. In some embodiments, the temperature at the target site is elevated to about 42°C to about 43°C within at most 30 minutes from the applying. In some embodiments, the temperature at the target site is elevated to about 42°C to about 43°C within 10 minutes or less after the applying. In some embodiments, the temperature at the target site is elevated to about 42°C to about 43°C within about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, or about 10 minutes after the applying.

[0038] In some embodiments, the temperature at the target site is maintained at about 42°C to about 43°C for at least about 15 minutes to at least about 120 minutes. In some embodiments, the temperature at the target site is maintained at 42°C to 43°C for at least 15 minutes, at least 20 minutes, at least 25 minutes, at least 30 minutes, at least 35 minutes, at least 40 minutes, at least 45 minutes, at least 50 minutes, at least 55 minutes, at least 60 minutes, at least 90 minutes, or at least 120 minutes. In some embodiments, the temperature at the target site is maintained at 42°C to 43°C for at least 20 minutes. In some embodiments, the temperature at the target site is maintained at about 42°C to about 43 °C for about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes.

[0039] In some embodiments, applying thermal energy to the target site of the subject comprises the application of one or more of ciliary body hyperthermia, magnetic hyperthermia, microwave hyperthermia, inductive diathermy, contact hyperthermia, ultrasonic diathermy, focused ultrasound, and deep hyperthermia. In some embodiments, applying thermal energy to the target site of the subject comprises use of a laser. In some embodiments, the laser is a 577 nm or a 532 nm laser. In some embodiments, the laser is set at a power level of about 25% to about 40%. In some embodiments, the laser is set at a power level of about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, or about 40%. In some embodiments, thelaser is set at a power level of about 30%. In some embodiments, the laser is applied to the target site for a pulse of about 1 second to about 15 seconds. In some embodiments, the laser is applied to the target site for a pulse of about 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 11 seconds, 12 seconds, 13 seconds, 14 seconds, or 15 seconds.

[0040] In some embodiments, upon applying thermal energy to the target site, the transcription of the therapeutic gene, the transactivator gene, the recombinase gene, or any combination thereof, increases by at least 1.1 -fold. In some embodiments, upon applying thermal energy to the target site, the levels of the therapeutic gene product, the transactivator gene product, the recombinase gene product, or any combination thereof, increases by at least 1.1- fold, wherein the gene product comprises RNA transcribed from the gene. In some embodiments, the gene product comprises an mRNA, a protein translated from the mRNA, or both.

[0041] In some embodiments, central retinal thickness (CRT), best-corrected visual acuity (BCVA) or both, are improved in the subject following the applying of thermal energy. In some embodiments, the method comprises applying thermal energy to the subject two or more times. In some embodiments, each of the two or more applications of thermal energy are 1 week apart, 2 weeks apart, 3 weeks apart, 4 weeks apart, 5 weeks apart, or more. In some embodiments, each of the two or more applications of thermal energy are applied when: (a) the best-corrected visual acuity (BCVA) of the subject decreases by at least 5 letters, and / or (b) the central retinal thickness CRT increases by at least 100 pM. In some embodiments, the decrease of (a) and the increase of (b) is relative to the BCVA and CRT (i) in a subject that has not been administered the composition; (ii) prior to administering the composition to the subject; (iii) prior to the first application of thermal energy to the subject, or (iv) following a prior application of the thermal energy to the subject.

[0042] In some embodiments, the retinal disease comprises age-related macular degeneration (AMD), non-infectious uveitis, Choroidemia, Leber congenital amaurosis (LCA), Leber hereditary optic neuropathy (LHON), Retinitis pigmentosa (RP), Achromatopsia, or Retinoscisis. In some embodiments, the AMD is wet or dry AMD.BRIEF DESCRIPTION OF THE DRAWINGS

[0043] FIG. 1 depicts a non-limiting exemplary schematic of methods and compositions disclosed herein.

[0044] FIG. 2 depicts a non-limiting exemplary schematic of activation and boost circuits disclosed herein.

[0045] FIG. 3 depicts 2D and 3D plots depicting data related to the duration and temperatures for activating the disclosed circuits.

[0046] FIG. 4 depicts exemplary diagrams of thermal circuits disclosed herein.

[0047] FIG. 5 depicts data related to the strength of different constitutive promoters that can be used in the compositions disclosed herein. FIG. 5 is original FIG. 2A from Fonseca, Joao Pedro, et al. "A toolkit for rapid modular construction of biological circuits in mammalian cells." ACS synthetic biology 8.11 (2019): 2593-2606. In some embodiments, different constitutive promoters can span 2-3 orders of magnitude in expression intensity.

[0048] FIG. 6 depicts a non-limiting exemplary schematic of the relationship between the length of CMV promoters and the level of expression.

[0049] FIGS. 7A-7D depict a non-limiting exemplary AAV circuit design (FIG. 7A), exemplary schematic (FIG. 7B), and data (FIGS. 7C-7D) related to studies thereof.DETAILED DESCRIPTION

[0050] In the following detailed description, reference is made to the accompanying drawings, which form a part hereof. In the drawings, similar symbols typically identify similar components, unless context dictates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not meant to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that the aspects of the present disclosure, as generally described herein, and illustrated in the Figures, can be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are explicitly contemplated herein and made part of the disclosure herein.

[0051] All patents, published patent applications, other publications, and sequences from GenBank, and other databases referred to herein are incorporated by reference in their entirety with respect to the related technology.

[0052] Disclosed herein include compositions for treating a retinal disease or disorder in a subject. In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product; wherein the first inducible promoter is capable of inducing transcription of the gene to generate the therapeutic gene product upon thermal stimulation.

[0053] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a transactivator gene, and a second promoter operably linked to a second polynucleotide comprising a therapeutic gene, wherein the first inducible promoter is capable of inducingtranscription of the transactivator gene to generate a transactivator transcript in the presence of thermal stimulation, wherein the transactivator transcript is capable of being translated to generate a transactivator; and wherein, in the presence of the transactivator and; optionally, a transactivator-binding compound, the second promoter is capable of inducing transcription of a therapeutic gene to generate a therapeutic gene product.

[0054] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter and a second promoter each operably linked to a first polynucleotide comprising a therapeutic gene and to a second polynucleotide comprising a transactivator gene, wherein the first inducible promoter is capable of inducing transcription of the therapeutic gene and the transactivator gene to generate a polycistronic transcript upon thermal stimulation, wherein, in the presence of the transactivator and a transactivator-binding compound, the second promoter is capable of inducing transcription of the therapeutic gene and the transactivator gene to generate a polycistronic transcript, and wherein the polycistronic transcript is capable of being translated to generate a transactivator and a therapeutic protein.

[0055] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a recombinase gene, wherein the first inducible promoter is capable of inducing transcription of the recombinase gene to generate a recombinase transcript upon thermal stimulation, and wherein the recombinase transcript is capable of being translated to generate a recombinase; a second promoter and a second polynucleotide comprising a therapeutic gene, wherein, in the absence of a recombination event, the second promoter and the second polynucleotide are not operably linked, wherein the recombinase is capable of catalyzing the recombination event, and wherein the second promoter and the second polynucleotide are operably linked after the recombination event such that the second promoter is capable of inducing transcription of the therapeutic gene to generate a therapeutic gene product.

[0056] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a leaky inducible promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product, wherein the leaky inducible promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the leaky inducible promoter is capable of boosting transcription of the gene, thereby driving a second level of expression of the therapeutic gene product upon thermal stimulation.

[0057] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a tandem promoter operably linked to a polynucleotide comprising a geneencoding a therapeutic gene product, wherein the tandem promoter comprises a first inducible promoter and a second constitutive promoter, wherein the second constitutive promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the first inducible promoter is capable of boosting transcription of the gene upon thermal stimulation, thereby driving a second level of expression of the therapeutic gene product.

[0058] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a gene encoding a therapeutic gene product and a second constitutive promoter operably linked to a second polynucleotide comprising the gene encoding a therapeutic gene product wherein the second constitutive promoter induces transcription of the gene encoding a therapeutic gene product from the second polynucleotide in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the first inducible promoter is capable of inducing the transcription of the therapeutic gene from the first polynucleotide upon thermal stimulation, thereby boosting the expression of the therapeutic gene product to a second level of expression.

[0059] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a bidirectional promoter comprising a first inducible promoter in a first orientation, and a second constitutive promoter in a second orientation, wherein the first inducible promoter is operably linked to a first polynucleotide comprising a first gene encoding a therapeutic gene product, and wherein the second constitutive promoter is operably linked to a second polynucleotide comprising a second gene, wherein the first and second orientations are opposite of each other; wherein the second constitutive promoter is capable of inducing transcription of the second gene in the absence of thermal stimulation; and wherein the first inducible promoter is capable of inducing transcription of the first gene upon thermal stimulation to generate the therapeutic gene product.

[0060] Provided herein are compositions for use in treating a retinal disease or disorder. Disclosed herein include methods of treating a retinal disease or disorder in a subject. In some embodiments, the method comprises: administering any of the compositions disclosed herein to the subject; and applying thermal energy to a target site of the subject sufficient to increase local temperature of the target site to an activating temperature, thereby inducing the expression of the therapeutic gene product.Definitions

[0061] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs. See, e.g. Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press (Cold Spring Harbor, NY 1989). For purposes of the present disclosure, the following terms are defined below.

[0062] As used herein, the terms “nucleic acid” and “polynucleotide” are interchangeable and refer to any nucleic acid, whether composed of phosphodiester linkages or modified linkages such as phosphotriester, phosphoramidate, siloxane, carbonate, carboxymethylester, acetamidate, carbamate, thioether, bridged phosphoramidate, bridged methylene phosphonate, bridged phosphoramidate, bridged phosphoramidate, bridged methylene phosphonate, phosphorothioate, methylphosphonate, phosphorodithioate, bridged phosphorothioate or sultone linkages, and combinations of such linkages. The terms “nucleic acid” and “polynucleotide” also specifically include nucleic acids composed of bases other than the five biologically occurring bases (adenine, guanine, thymine, cytosine and uracil).

[0063] The term “vector” as used herein, can refer to a vehicle for carrying or transferring a nucleic acid. Non-limiting examples of vectors include plasmids and viruses (for example, AAV viruses).

[0064] The term “construct,” as used herein, refers to a recombinant nucleic acid that has been generated for the purpose of the expression of a specific nucleotide sequence(s), or that is to be used in the construction of other recombinant nucleotide sequences.

[0065] As used herein, the term “plasmid” refers to a nucleic acid that can be used to replicate recombinant DNA sequences within a host organism. The sequence can be a double stranded DNA.

[0066] The term “element” refers to a separate or distinct part of something, for example, a nucleic acid sequence with a separate function within a longer nucleic acid sequence. The term “regulatory element” and “expression control element” are used interchangeably herein and refer to nucleic acid molecules that can influence the expression of an operably linked coding sequence in a particular host organism. These terms are used broadly to and cover all elements that promote or regulate transcription, including promoters, core elements required for basic interaction of RNA polymerase and transcription factors, upstream elements, enhancers, and response elements (see, e.g., Lewin, “Genes V” (Oxford University Press, Oxford) pages 847- 873). Exemplary regulatory elements in prokaryotes include promoters, operator sequences and aribosome binding sites. Regulatory elements that are used in eukaryotic cells can include, without limitation, transcriptional and translational control sequences, such as promoters, enhancers, splicing signals, polyadenylation signals, terminators, protein degradation signals, internal ribosome-entry element (IRES), 2A sequences, and the like, that provide for and / or regulate expression of a coding sequence and / or production of an encoded polypeptide in a host cell.

[0067] As used herein, the term “promoter” is a nucleotide sequence that permits binding of RNA polymerase and directs the transcription of a gene. Typically, a promoter is located in the 5’ non-coding region of a gene, proximal to the transcriptional start site of the gene. Sequence elements within promoters that function in the initiation of transcription are often characterized by consensus nucleotide sequences. Examples of promoters include, but are not limited to, promoters from bacteria, yeast, plants, viruses, and mammals (including humans). A promoter can be inducible, repressible, and / or constitutive. Inducible promoters initiate increased levels of transcription from DNA under their control in response to some change in environmental conditions, such as a change in temperature. In some embodiments, the inducible promoter is a “heat-shock promoter” that is activated in response to heat (e.g., thermal energy). Constitutive promoters can have a first or basal level of expression, e.g., in most cells types and most developmental timepoints, e.g., are “always on” promoters.

[0068] As used herein, the term “enhancer” refers to a type of regulatory element that can increase the efficiency of transcription, regardless of the distance or orientation of the enhancer relative to the start site of transcription.

[0069] As used herein, the term “operably linked” can be used to describe the connection between regulatory elements and a gene or its coding region. Typically, gene expression is placed under the control of one or more regulatory elements, for example, without limitation, constitutive or inducible promoters, tissue-specific regulatory elements, and enhancers. A gene or coding region is said to be “operably linked to” or “operatively linked to” or “operably associated with” the regulatory elements, meaning that the gene or coding region is controlled or influenced by the regulatory element. For instance, a promoter is operably linked to a coding sequence if the promoter effects transcription or expression of the coding sequence.

[0070] The term “construct,” as used herein, refers to a recombinant nucleic acid that has been generated for the purpose of the expression of a specific nucleotide sequence(s), or that is to be used in the construction of other recombinant nucleotide sequences.

[0071] As used herein, a “subject” refers to an animal that is the object of treatment, observation or experiment. “Animal” includes cold- and warm-blooded vertebrates andinvertebrates such as fish, shellfish, reptiles, and in particular, mammals. “Mammal,” as used herein, refers to an individual belonging to the class Mammalia and includes, but not limited to, humans, domestic and farm animals, zoo animals, sports and pet animals. Non-limiting examples of mammals include mice; rats; rabbits; guinea pigs; dogs; cats; sheep; goats; cows; horses; primates, such as monkeys, chimpanzees and apes, and, in particular, humans. In some embodiments, the mammal is a human. However, in some embodiments, the mammal is not a human.

[0072] As used herein, the term “treatment” refers to an intervention made in response to a disease, disorder or physiological condition manifested by a patient. The aim of treatment may include, but is not limited to, one or more of the alleviation or prevention of symptoms, slowing or stopping the progression or worsening of a disease, disorder, or condition and the remission of the disease, disorder or condition. The term “treat” and “treatment” includes, for example, therapeutic treatments, prophylactic treatments, and applications in which one reduces the risk that a subject will develop a disorder or other risk factor. Treatment does not require the complete curing of a disorder and encompasses embodiments in which one reduces symptoms or underlying risk factors. In some embodiments, “treatment” refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already affected by a disease or disorder or undesired physiological condition as well as those in which the disease or disorder or undesired physiological condition is to be prevented. As used herein, the term “prevention” refers to any activity that reduces the burden of the individual later expressing those symptoms. This can take place at primary, secondary and / or tertiary prevention levels, wherein: a) primary prevention avoids the development of symptoms / disorder / condition; b) secondary prevention activities are aimed at early stages of the condi tion / disorder / symptom treatment, thereby increasing opportunities for interventions to prevent progression of the condition / disorder / symptom and emergence of symptoms; and c) tertiary prevention reduces the negative impact of an already established condition / disorder / symptom by, for example, restoring function and / or reducing any condition / disorder / symptom or related complications. The term “prevent” does not require the 100% elimination of the possibility of an event. Rather, it denotes that the likelihood of the occurrence of the event has been reduced in the presence of the compound or method.

[0073] As used herein, the term “effective amount” refers to an amount sufficient to effect beneficial or desirable biological and / or clinical results.

[0074] “Pharmaceutically acceptable” carriers are ones which are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. “Pharmaceuticallyacceptable” carriers can be, but not limited to, organic or inorganic, solid or liquid excipients which is suitable for the selected mode of application such as oral application or injection, and administered in the form of a conventional pharmaceutical preparation, such as solid such as tablets, granules, powders, capsules, and liquid such as solution, emulsion, suspension and the like. Often the physiologically acceptable carrier is an aqueous pH buffered solution such as phosphate buffer or citrate buffer. The physiologically acceptable carrier may also comprise one or more of the following: antioxidants including ascorbic acid, low molecular weight (less than about 10 residues) polypeptides, proteins, such as serum albumin, gelatin, immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone, amino acids, carbohydrates including glucose, mannose, or dextrins, chelating agents such as EDTA, sugar alcohols such as mannitol or sorbitol, salt-forming counterions such as sodium, and nonionic surfactants such as Tween™, polyethylene glycol (PEG), and Pluronics™. Auxiliary, stabilizer, emulsifier, lubricant, binder, pH adjustor controller, isotonic agent and other conventional additives may also be added to the carriers.

[0075] The term “antibody fragment” shall be given its ordinary meaning, and shall also refers to at least one portion of an antibody, that retains the ability to specifically interact with (e.g., by binding, steric hindrance, stabilizing / destabilizing, spatial distribution) an epitope of an antigen. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, scFv antibody fragments, disulfide-linked Fvs (sdFv), a Fd fragment consisting of the VH and CHI domains, linear antibodies, single domain antibodies such as sdAb (either VL or VH), camelid VHH domains, multi-specific antibodies formed from antibody fragments such as a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region, and an isolated CDR or other epitope binding fragments of an antibody. An antigen binding fragment can also be incorporated into single domain antibodies, maxibodies, minibodies, nanobodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv (see, e.g., Hollinger and Hudson, Nature Biotechnology 23 : 1126-1136, 2005). Antigen binding fragments can also be grafted into scaffolds based on polypeptides such as a fibronectin type III (Fn3) (see U.S. Pat. No. 6,703,199, which describes fibronectin polypeptide minibodies).

[0076] Disclosed herein are methods and compositions for using pHSPs (heat shock protein promoters) to activate or boost the production of AAV-delivered biologies. The presently disclosed methods and compositions advantageously allow for activation of expression of therapeutic molecules under control of pHSP for treatment of a disease or disorder (e.g., a retinal disease or disorder) on a schedule or in response to flare-ups. The presently disclosed methods and compositions allows the therapy to remain off or low, when not needed or causing side-effects. Also disclosed herein are boosting paradigms, in which therapeutic molecule expression is regulated by, e.g., a constitutive promoter and pHSP, to allow boosting when necessary above a baseline level. The present disclosure meets several unmet needs, including the inability to readminister AAV to boost therapy and allows response to flares or other transient needs.

[0077] Previous AAV-therapy for retinal diseases (e.g., age-related macular degeneration (AMD)) have established the retina as a site for local biologic treatment using AAV. For example, in wet AMD, atypical blood vessels appear around the macula and retina. These vessels can leak proteins or lipids, and they may also cause scarring. Anti -angiogenic therapies can be effective in treating diseases such as AMD.

[0078] In some embodiments, the therapeutic gene product is an inhibitor of VEGF. Vascular endothelial growth factor (VEGF) is the main protein that causes new blood vessels to form in the macula in wet AMD. VEGF inhibitors reduce leakage from blood vessels, prevent their growth, decrease swelling of the retina, reduce vision loss, and improve vision in wet AMD. Several AAV-based therapies delivering VEGFi to the retina have been approved, however, these typically require repeated intravitreal injection, which carries risk and is aversive to patients (e.g., every 1-3 months). A concern of current AAV strategies is that insufficient drug production compromises efficacy; but too much always-on production may lead to side-effects (e.g., macular atrophy with VEGFi). The presently disclosed methods and compositions address this concern with activation or boosting, either on a scheduled, or as needed using thermal activation.

[0079] Disclosed herein include compositions for treating a retinal disease or disorder in a subject. In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product; wherein the first inducible promoter is capable of inducing transcription of the gene to generate the therapeutic gene product upon thermal stimulation.

[0080] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a transactivator gene, and a second promoter operably linked to a second polynucleotide comprising a therapeutic gene, wherein the first inducible promoter is capable of inducing transcription of the transactivator gene to generate a transactivator transcript in the presence of thermal stimulation, wherein the transactivator transcript is capable of being translated to generate a transactivator; and wherein, in the presence of the transactivator and; optionally, a transactivator-binding compound, the second promoter is capable of inducing transcription of a therapeutic gene to generate a therapeutic gene product.

[0081] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter and a second promoter each operably linked to a first polynucleotide comprising a therapeutic gene and to a second polynucleotide comprising a transactivator gene, wherein the first inducible promoter is capable of inducing transcription of the therapeutic gene and the transactivator gene to generate a polycistronic transcript upon thermal stimulation, wherein, in the presence of the transactivator and a transactivator-binding compound, the second promoter is capable of inducing transcription of the therapeutic gene and the transactivator gene to generate a polycistronic transcript, and wherein the polycistronic transcript is capable of being translated to generate a transactivator and a therapeutic protein.

[0082] The second promoter can comprise one or more copies of a transactivator recognition sequence the transactivator is capable of binding to induce transcription. In some embodiments, the transactivator is incapable of binding the transactivator recognition sequence in the absence of the transactivator-binding compound. The one or more copies of a transactivator recognition sequence can comprise one or more copies of a tet operator (TetO). The transactivator can comprise reverse tetracycline-controlled transactivator (rtTA). The transactivator can comprise tetracycline-controlled transactivator (tTA). The transactivator-binding compound can comprise tetracycline, doxycycline or a derivative thereof. The first polynucleotide and the second polynucleotide can be operably linked to a tandem gene expression element. The tandem gene expression element can be an internal ribosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof.

[0083] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a recombinase gene, wherein the first inducible promoter is capable of inducing transcription of the recombinase gene to generate a recombinase transcript upon thermal stimulation, and wherein the recombinase transcript is capable of being translated to generate a recombinase; a second promoter and a second polynucleotide comprising a therapeutic gene, wherein, in the absence of a recombination event, the second promoter and the second polynucleotide are not operably linked, wherein the recombinase is capable of catalyzing the recombination event, and wherein the second promoter and the second polynucleotide are operably linked after the recombination event such that the second promoter is capable of inducing transcription of the therapeutic gene to generate a therapeutic gene product. The recombination event can comprise removal of a sequence flanked by recombinase target sites or an inversion of a sequence flanked by recombinase target sites. The second polynucleotide can be flanked by recombinase targetsites. Prior to the recombination event, the sequence of the therapeutic gene can be inverted relative to the promoter. The composition can comprise at least one stop cassette situated between the second promoter and the therapeutic gene, wherein the stop cassette comprises one or more stop sequences, and wherein the one or more stop cassettes are flanked by recombinase target sites. The therapeutic gene product can be an RNA transcript and / or protein. The transcript can be an mRNA capable of being translated to generate the protein. The at least one stop cassette can be configured to prevent transcription of the therapeutic gene and / or translation of the therapeutic transcript. The one or more stop sequences can comprise a polyadenylation signal, a stop codon, a frame-shifting mutation, or any combination thereof. The second promoter can comprise a ubiquitous or tissue-specific promoter. In some embodiments, the second promoter comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focus-forming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl 1 promoters from vaccinia virus, an elongation factor 1 -alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3 -phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P-kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CBH promoter, any variant thereof, or any combination thereof. The recombinase can be Cre, Dre, Flp, KD, B2, B3, , HK022, HP1, y6, ParA, Tn3, Gin, C31, Bxbl, R4, derivatives thereof, or any combination thereof. The recombinase can be a Flp recombinase and the recombinase target sites can be FRT sites or the recombinase can be a Cre recombinase and the recombinase target sites can be loxP sites.Inducible promoters and thermal circuits

[0084] There are provided, in some embodiments, promoters capable of inducing transcription upon thermal stimulation (e.g., inducible promoters). The first inducible promoter can comprise or can be derived from a mammalian heat shock promoter (HSP) or a C. elegans HSP. The mammalian HSP can be a human HSP or mouse HSP. The first inducible promoter can comprise a nucleotide sequence that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of SEQ ID NOs: 1-34. The first inducible promoter can comprise one or more AP-1 sites. In some embodiments, the first inducible promoter does not comprise an AP-1site. The first inducible promoter can comprise a bidirectional promoter and / or a minimal bidirectional promoter. The first inducible promoter can comprise one or more heat shock element (HSE) binding sites (e.g., seven HSE binding sites). In some embodiments, the first inducible promoter does not comprise a human transcription factor binding site other than one or more HSE binding sites. In some embodiments, the first inducible promoter comprises one or more of a TATA box, GC-Box, CAAT signal, and AP-1 site. Nucleic acids provided herein can comprise a portion of a promoter, an enhancer, positive or negative cis-acting sequences, inducible or repressible control element, 5’ UTR sequences that are upstream of a gene, or any combination thereof. A disclosed promoter (e.g., first inducible promoter) can comprise 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 HSE binding sites. The inducible promoter can comprise a promoter sequence shown in Table 1.

[0085] A disclosed promoter (e.g., first inducible promoter) can be derived from the heat shock promoter (HSP) of one or more species selected from the group comprising: Arabidopsis thaliana; Aspergillus nidulans; Bombyx mori; Candida albicans; Caenorhabditis elegans; Chlamydomonas rheinhardtii; Cricetulus griseus; Cyanophora paradoxa; Cylindrotheca fusiformis; Danio rerio; Dictyostelium discoideum; Drosophila melanogaster; Drosophila yakuba; Gallus gallus; Homo Sapiens; Leishmania chagasi; Leishmania major; Loligo pealii; Lymantria dispar; Monodelphis domestica; Morone saxatilis; Mus musculus; Nectria haematococca; Neurospora crassa; Nicotiana tabacum; Oryza sativa; Paracentrotus lividus; Plasmodium falciparum; Rattus norvegicus; Saccharomyces cerevisiae; Schizosaccharomyces pombe; Solanum tuberosum; Strongylocentrotus purpuratus; Syncephalastrum racemosum; Tetrahymena thermophila; Trypanosoma brucei; Ustilago maydis; Volvox carleri: and Xenopus laevis.

[0086] The length of the promoters provided herein (e.g., first inducible promoter) can vary. In some embodiments, a disclosed promoter is, or is about, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 110, 120, 128, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270,280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460,470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650,660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840,850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850,1900, 1950, 1100, 1150, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500,1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950, 2000, 2050, 2100, 2150, 2200, 2250,2300, 2350, 2400, 2450, 2500, 2550, 2600, 2650, 2700, 2750, 2800, 2850, 2900, 2950, 3000, or4000, or a number or a range between any two of these values, nucleotides in length. In some embodiments, a disclosed promoter is at least, or is at most, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 110, 120, 128, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270,280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460,470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650,660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840,850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850,1900, 1950, 1100, 1150, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500,1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950, 2000, 2050, 2100, 2150, 2200, 2250,2300, 2350, 2400, 2450, 2500, 2550, 2600, 2650, 2700, 2750, 2800, 2850, 2900, 2950, 3000, or4000, nucleotides in length.

[0087] In some embodiments, the sequence identity between a disclosed promoter (e.g., first inducible promoter) and the sequence of any one of SEQ ID NOs: 1-34 can be, or be about, 0.000000001%, 0.00000001%, 0.0000001%, 0.000001%, 0.00001%, 0.0001%, 0.001%, 0.01%, 0.1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values. In some embodiments, the sequence identity between a disclosed promoter and the sequence of any one of SEQ ID NOs: 1-34 can be at least, or at most, 0.000000001%, 0.00000001%, 0.0000001%, 0.000001%, 0.00001%, 0.0001%, 0.001%, 0.01%, 0.1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0088] A disclosed promoter (e.g., first inducible promoter) can comprise at least about 20 consecutive nucleotides (e.g., about 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 60 nt, 70 nt, 80 nt, 90 nt, 100 nt, 110 nt, 120 nt, 128 nt, 130 nt, 140 nt, 150 nt, 160 nt, 170 nt, 180 nt, 190 nt, 200nt, 210 nt, 220 nt, 230 nt, 240 nt, 250 nt, 260 nt, 270 nt, 280 nt, 290 nt, 300 nt, 310 nt, 320 nt, 330 nt, 340 nt, 350 nt, 360 nt, 370 nt, 380 nt, 390 nt, 400 nt, 410 nt, 420 nt, 430 nt, 440 nt, 450 nt, 460 nt, 470 nt, 480 nt, 490 nt, 500 nt, 510 nt, 520 nt, 530 nt, 540 nt, 550 nt, 560 nt, 570 nt, 580 nt, 590 nt, 600 nt, 610 nt, 620 nt, 630 nt, 640 nt, 650 nt, 660 nt, 670 nt, 680 nt, 690 nt, 700 nt, 710 nt, 720 nt, 730 nt, 740 nt, 750 nt, 760 nt, 770 nt, 780 nt, 790 nt, 800 nt, 810 nt, 820 nt, 830 nt, 840 nt, 850 nt, 860 nt, 870 nt, 880 nt, 890 nt, 900 nt, 910 nt, 920 nt, 930 nt, 940 nt, 950 nt, 960 nt, 970 nt, 980 nt, 990 nt, 1000 nt, or a number or a range between any two of these values) of a sequences described by SEQ ID NOs: 1-34.

[0089] The first inducible promoter can comprise a core promoter and at least one heat shock element (HSE). In some embodiments, the core promoter comprises the heat shock protein transcription start site of a gene selected from the group consisting of HSPA1A, HSPH1, HSPB1, HSPA6, and YB.

[0090] The core promoter can comprise or consist of the sequence of any one of SEQ ID NOs: 9-12 or a sequence having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity, or a number or a range between any two of these values) to any one of SEQ ID NOs: 9-12. In some embodiments, the core promoter comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 9-12.

[0091] The first inducible promoter can comprise seven HSEs. Each of the at least one HSE can comprise the sequence of 5’- nGAAnnTTCnnGAAn-3’. The at least one HSE can comprise or consist of the sequence of any one of SEQ ID NOs: 1-8 or a sequence having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity, or a number or a range between any two of these values) to any one of SEQ ID NOs: 1-8 or a sequence differing by one or two nucleotide mismatches relative to any one of SEQ ID NOs: 1-8. In some embodiments, the at least one HSE comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 1-8.

[0092] The first inducible promoter can comprise or consist of the sequence of any one of SEQ ID NOs: 13-20 or a sequence having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity, or a number or a range between any two of these values) to any one of SEQ ID NOs: 13-20. In some embodiments, the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 13-20. The first inducible promoter can comprise or consist of the sequence of any one of SEQ ID NOs: 21-34 or a sequence having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%identity, or a number or a range between any two of these values) to any one of SEQ ID NOs: 21-34. In some embodiments, the first inducible promoter comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 21-34. Table 1 below displays exemplary heat inducible promoter sequences disclosed herein. Certain embodiments of the thermal circuits described herein are also described in PCT publications WO2022 / 272102 and WO2021 / 211776, the contents of which are hereby incorporated by reference in their entireties.TABLE 1 : EXEMPLARY INDUCIBLE PROMOTER SEQUENCES

[0093] In some embodiments, thermal stimulation comprises heating to an activating temperature, wherein the activating temperature is greater than 37°C. The activating temperature can comprise a temperature of at least 37°C to at most 70°C. The activating temperature can be about 37.0°C, 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, about 46.0°C, about 46.5°C, about 47.0°C, about 47.5°C, about 48.0°C, about 48.5 °C, about 49.0°C, about 49.5°C, about 50.0°C, about 50.5°C, about 51.0°C, about 51.5°C, about 52.0°C, about 52.5°C, about 53.0°C, about 53.5°C, about 54.0°C, about 54.5°C, about 55.0°C, about 55.5°C, about 56.0°C, about 56.5°C, about 57.0°C, about 57.5°C, about 58.0°C, about 58.5°C, about 59.0°C, about 59.5°C, about 60.0°C, about 60.5°C, about 61.0°C, about 61.5°C, about 62.0°C, about 62.5°C, about 63.0°C, about 63.5°C, about 64.0°C, about 64.5°C, about 65.0°C, about 65.5°C, about 66.0°C, about 66.5°C, about 67.0°C, about 67.3°C, about 67.5°C, about 68.0°C, about 68.5°C, about 69.0°C, about 69.5°C, or about 70.0°C. The activating temperature can be about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, or about 46.0°C.

[0094] The activating temperature can comprise a temperature of about 42°C to about 43°C. The activating temperature can be about 42.0°C, about 42.1°C, about 42.2°C, about 42.3°C, about 42.4°C, about 42.5°C, about 42.6°C, about 42.7°C, about 42.8°C, about 42.9°C, or about 43.0°C.Expression Levels and Tuning

[0095] In some embodiments, in the absence of thermal stimulation, the therapeutic gene product reaches unstimulated steady state levels in a target region (e.g., retina). Unstimulated steady state levels of the therapeutic gene product can be insufficient to exert a phenotypic effect and / or therapeutic effect. In some embodiments, upon thermal stimulation transcription of the therapeutic gene product, transactivator gene, oscillator gene, and / or recombinase gene from the first inducible promoter is increased by at least 1.1-fold fold (e.g., 1.1-fold, 1.3-fold, 1.5-fold, 1.7-fold, 1.9-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, or a number or a range between any of these values). In some embodiments, increasing transactivator-binding compound concentration increases stimulated steady state levels.

[0096] In some embodiments, the steady-state levels of the therapeutic gene transcript, the steady-state levels of transactivator transcript, the steady-state levels of recombinase transcript, the steady-state levels of oscillator transcript, and / or the steady-state levels of the polycistronic transcript are at least 1.1-fold (e.g., 1.1-fold, 1.3-fold, 1.5-fold, 1.7-fold, 1.9-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, or a number or a range between any of these values) higher upon thermal stimulation. In some embodiments, upon thermal stimulation, the therapeutic gene product reaches stimulated steady state therapeutic gene product levels in a target region. In some embodiments, the therapeutic gene product does not return to unstimulated steady state therapeutic gene product levels.

[0097] Stimulated steady state therapeutic gene product levels can be at least 1.1 -fold fold (e.g., 1.1-fold, 1.3-fold, 1.5-fold, 1.7-fold, 1.9-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, or a number or a range between any of these values) higher than unstimulated steady state therapeutic gene product levels.

[0098] In some embodiments, after a first duration of time, the therapeutic gene product levels returns to unstimulated steady state therapeutic gene product levels from stimulated steady state therapeutic gene product levels, wherein the first duration of time is about 250 hours, about 200 hours, about 150 hours, about 96 hours, about 48 hours, about 44 hours, about 40 hours, about 35 hours, about 30 hours, about 25 hours, 20 hours, 15 hours, 10 hours, about 8 hours, about 8 hours, 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, about 10 minutes, about 5 minutes, or a number or a range between any two of these values. In some embodiments, the first duration of time is greater than 250 hours.

[0099] In some embodiments, the nucleic acid comprises: a transcript stabilization element. The transcript stabilization element can comprise woodchuck hepatitis post-translational regulatory element (WPRE), bovine growth hormone polyadenylation (bGH-polyA) signalsequence, human growth hormone polyadenylation (hGH-polyA) signal sequence, or any combination thereof. The therapeutic gene can comprise a 5’UTR and / or a 3’UTR. The transactivator gene can comprise a 5’UTR and / or a 3’UTR. The recombinase gene can comprise a 5’UTR and / or a 3’UTR. The oscillator gene can comprise a 5’UTR and / or a 3’UTR. The 5’ UTR can comprise a Kozak sequence. In some embodiments, stimulated steady state therapeutic gene product levels, unstimulated steady state therapeutic gene product levels, the lower tuned threshold, and / or the upper tuned threshold can be tuned by adjusting the presence and / or sequence of the Kozak sequence. The 5’ UTR can comprise one or more micro open reading frames. In some embodiments, stimulated steady state therapeutic gene product levels, unstimulated steady state therapeutic gene product levels, the lower tuned threshold, and / or the upper tuned threshold can be tuned by adjusting the presence and / or sequence of the one or more micro open reading frames.Constitutive promoters

[0100] In some embodiments, a nucleotide sequence encoding a gene product of interest is operably linked to a constitutive promoter. In other embodiments, a nucleotide sequence encoding a gene product of interest is operably linked to an inducible promoter. In some instances, a nucleotide sequence encoding a gene product of interest is operably linked to a tissue- specific or cell type- specific regulatory element. For example, in some instances, a nucleotide sequence encoding a gene product of interest is operably linked to a photoreceptorspecific regulatory element (e.g., a photoreceptor-specific promoter), e.g., a regulatory element that confers selective expression of the operably linked gene in a photoreceptor cell. Suitable photoreceptor- specific regulatory elements include, e.g., a rhodopsin promoter; a rhodopsin kinase promoter (Young et al. (2003) Ophthalmol. Vis. Sci. 44:4076); a beta phosphodiesterase gene promoter (Nicoud et al. (2007) J. Gene Med. 9: 1015); a retinitis pigmentosa gene promoter (Nicoud et al. (2007) supra); an interphotoreceptor retinoid-binding protein (IRBP) gene enhancer (Nicoud et al. (2007) supra); an IRBP gene promoter (Yokoyama et al. (1992) Exp Eye Res. 55:225).

[0101] In some embodiments, the constitutive promoter (e.g., the second constitutive promoter) comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focus-forming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl 1 promoters from vaccinia virus, an elongation factor 1-alpha(EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3 -phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P-kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CBH promoter, or any combination thereof.

[0102] The first inducible promoter and the second constitutive promoter can be in the same or opposite orientations. In some embodiments, any two of the first inducible promoter, the second promoter, the leaky inducible promoter, the tandem promoter, the second constitutive promoter, and the bidirectional promoter are in opposite orientations or the same orientation relative to each other.

[0103] As shown in, e.g., FIG. 5-FIG. 6, different constitutive promoters can induce different levels of basal (e.g., a first level) of expression. The skilled artisan will be able to assess and choose a suitable constitutive promoter with a desired basal expression level.Boost paradigms

[0104] In some embodiments, methods and compositions are disclosed for providing a therapeutic boost. In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a leaky inducible promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product, wherein the leaky inducible promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the leaky inducible promoter is capable of boosting transcription of the gene, thereby driving a second level of expression of the therapeutic gene product upon thermal stimulation.

[0105] The leaky inducible promoter can comprise an HSPB, HSPB’2, HSP A / A, or HSP A / B promoter. The leaky inducible promoter can comprise the sequence of any one of SEQ ID NOs: 21-24 and 28-29 or a sequence having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity, or a number or a range between any two of these values) to any one of SEQ ID NOs: 21-24 and 28-29. In some embodiments, the leaky inducible promoter comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 21-24 and 28-29.

[0106] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a tandem promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product, wherein the tandem promoter comprises a first induciblepromoter and a second constitutive promoter, wherein the second constitutive promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the first inducible promoter is capable of boosting transcription of the gene upon thermal stimulation, thereby driving a second level of expression of the therapeutic gene product. The tandem promoter can comprise, from 5’ to 3’: the first inducible promoter then the second constitutive promoter. The tandem promoter can comprise, from 5’ to 3’ : the second constitutive promoter then the first inducible promoter.

[0107] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a gene encoding a therapeutic gene product and a second constitutive promoter operably linked to a second polynucleotide comprising the gene encoding a therapeutic gene product wherein the second constitutive promoter induces transcription of the gene encoding a therapeutic gene product from the second polynucleotide in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the first inducible promoter is capable of inducing the transcription of the therapeutic gene from the first polynucleotide upon thermal stimulation, thereby boosting the expression of the therapeutic gene product to a second level of expression.

[0108] The nucleic acid can comprise, from 5’ to 3’: the first inducible promoter, the first polynucleotide, the second constitutive promoter, and the second polynucleotide. The nucleic acid can comprise, from 5’ to 3’: the second constitutive promoter, the second polynucleotide, the first inducible promoter, and the first polynucleotide. The second level of expression can be at least 1.1 -fold higher (e.g., 1.1-fold, 1.3-fold, 1.5-fold, 1.7-fold, 1.9-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, or a number or a range between any of these values, or more) than the first level of expression. In some embodiments, after a first duration of time, the therapeutic gene product returns to the first level of expression from the second level of expression, wherein the first duration of time is about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks or more.

[0109] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a first gene encoding a therapeutic gene product and a second constitutive promoter operably linked to the first polynucleotide comprising the first gene encoding a therapeutic gene product and a second polynucleotide comprising a second gene, wherein the second constitutive promoter induces transcription of the first and second genes in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product and thesecond gene product; and wherein the first inducible promoter is capable of boosting transcription of the first gene upon thermal stimulation, thereby driving a second level of expression of the therapeutic gene product. In some embodiments, the nucleic acid comprises, from 5’ to 3’: the second constitutive promoter, the second polynucleotide, the first inducible promoter, and the first polynucleotide.

[0110] In some embodiments, the composition comprises: a viral vector comprising a nucleic acid comprising a bidirectional promoter comprising a first inducible promoter in a first orientation, and a second constitutive promoter in a second orientation, wherein the first inducible promoter is operably linked to a first polynucleotide comprising a first gene encoding a therapeutic gene product, and wherein the second constitutive promoter is operably linked to a second polynucleotide comprising a second gene, wherein the first and second orientations are opposite of each other; wherein the second constitutive promoter is capable of inducing transcription of the second gene in the absence of thermal stimulation; and wherein the first inducible promoter is capable of inducing transcription of the first gene upon thermal stimulation to generate the therapeutic gene product.[oni] In some embodiments, the second gene encodes a therapeutic gene product. The therapeutic gene product can be capable of inhibiting vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6). In some embodiments, the therapeutic gene product is selected from the group consisting of RPE65, Complement Factor 1 (CF1), CD59, FMS Related Receptor Tyrosine Kinase 1 (FLT-1), Rab escort protein 1 (REP1), NADH hydrogenase 4 (ND4), MER proto-oncogene tyrosine kinase (MERTK), X-linked retinitis pigmentosa GTPase regulator (RPGR), or Retinoschisin (RSI). The second constitutive promoter can comprise a ubiquitous or tissue-specific promoter. In some embodiments, the second constitutive promoter comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focus-forming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RS V) LTR, an RS V promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl 1 promoters from vaccinia virus, an elongation factor 1 -alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P-kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CB A) promoter, a CAG promoter, a CBH promoter, or any combination thereof.

[0112] The first inducible promoter can comprise a core promoter and at least one heat shock element (HSE). In some embodiments, the core promoter comprises the heat shock protein transcription start site of a gene selected from the group consisting of HSPA1A, HSPH1, HSPB1, HSPA6, and YB.

[0113] The core promoter can comprise the sequence of any one of SEQ ID NOs: 9-12 or a sequence having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity, or a number or a range between any two of these values) to any one of SEQ ID NOs: 9-12. In some embodiments, the core promoter comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 9-12.

[0114] The first inducible promoter can comprise seven HSEs. Each of the at least one HSE can comprise the sequence of 5’- nGAAnnTTCnnGAAn-3’. The at least one HSE can comprise the sequence of any one of SEQ ID NOs: 1-8 or a sequence having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity, or a number or a range between any two of these values) to any one of SEQ ID NOs: 1- 8 or a sequence differing by one or two nucleotide mismatches relative to any one of SEQ ID NOs: 1-8. In some embodiments, the at least one HSE comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 1-8. The first inducible promoter can comprise the sequence of any one of SEQ ID NOs: 13-20 or a sequence having at least 85% identity to any one of SEQ ID NOs: 13-20. In some embodiments, the first inducible promoter comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 13-20. The first inducible promoter can comprise the sequence of any one of SEQ ID NOs: 21-34 or a sequence having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity, or a number or a range between any two of these values) to any one of SEQ ID NOs: 21-34. In some embodiments, the first inducible promoter comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 21-34. The first inducible promoter and the second constitutive promoter can be in the same or opposite orientations. In some embodiments, any two of the first inducible promoter, the second promoter, the leaky inducible promoter, the tandem promoter, the second constitutive promoter, and the bidirectional promoter are in opposite orientations or the same orientation relative to each other.

[0115] The therapeutic gene product can comprise an siRNA, an shRNA, a miRNA, or a protein. The therapeutic gene product can be capable of inhibiting vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6). The therapeutic gene product can comprise an antibody or fragment thereof capable of binding tovascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6). The therapeutic gene product can comprise RPE65, Complement Factor 1 (CF1), CD59, FMS Related Receptor Tyrosine Kinase 1 (FLT-1), Rab escort protein 1 (REP1), NADH hydrogenase 4 (ND4), MER proto-oncogene tyrosine kinase (MERTK), X-linked retinitis pigmentosa GTPase regulator (RPGR), or Retinoschisin (RSI).Viral Vectors

[0116] There are provided, in some embodiments, viral vector compositions. The viral vector (e.g., a single viral vector, a first viral vector, a second viral vector, a primary viral vector, a secondary viral vector, a tertiary viral vector) can be, can comprise, or can be derived from, an AAV vector, a lentivirus vector, a retrovirus vector, an adenovirus vector, a herpesvirus vector, a herpes simplex virus vector, a cytomegalovirus vector, a vaccinia virus vector, a MVA vector, a baculovirus vector, a vesicular stomatitis virus vector, a human papillomavirus vector, an avipox virus vector, a Sindbis virus vector, a VEE vector, a Measles virus vector, an influenza virus vector, a hepatitis B virus vector, an integration-deficient lentivirus (IDLV) vector derivatives thereof, or any combination thereof. The viral vector can be capable of integrating into a mammalian cell genome.

[0117] In some embodiments, the viral vector is a recombinant lentiviral vector. In some embodiments, the viral particle is a lentiviral particle. Lentiviruses are positive-sense, ssRNA retroviruses with a genome of approximately 10 kb. Lentiviruses are known to integrate into the genome of dividing and non-dividing cells. Lentiviral particles may be produced, for example, by transfecting multiple plasmids (typically the lentiviral genome and the genes required for replication and / or packaging are separated to prevent viral replication) into a packaging cell line, which packages the modified lentiviral genome into lentiviral particles. In some embodiments, a lentiviral particle may refer to a first generation vector that lacks the envelope protein. In some embodiments, a lentiviral particle may refer to a second generation vector that lacks all genes except the gag / pol and tat / rev regions. In some embodiments, a lentiviral particle may refer to a third generation vector that only contains the endogenous rev, gag, and pol genes and has a chimeric LTR for transduction without the tat gene (see Dull. T. et al. (1998) J. Virol. 72:8463- 71). For further description, see Durand, S. and Cimarelli, A. (2011) Viruses 3 : 132-59.

[0118] Use of any lentiviral vector is considered within the scope of the disclosed compositions and methods. In some embodiments, the lentiviral vector is derived from a lentivirus including, without limitation, human immunodeficiency virus-1 (HIV-1), human immunodeficiency virus-2 (HIV-2), simian immunodeficiency virus (SIV), felineimmunodeficiency virus (FIV), equine infectious anemia virus (EIAV), bovine immunodeficiency virus (BIV), Jembrana disease virus (JDV), visna virus (VV), and caprine arthritis encephalitis virus (CAEV).

[0119] In some embodiments, the viral vector is encapsidated in a viral particle. In some embodiments, the viral particle is a recombinant lentiviral particle encapsidating a recombinant lentiviral vector. In some embodiments, the recombinant viral particles comprise a lentivirus vector in combination with one or more foreign viral capsid proteins. Such combinations may be referred to as pseudotyped recombinant lentiviral particles. In some embodiments, foreign viral capsid proteins used in pseudotyped recombinant lentiviral particles are derived from a foreign virus. In some embodiments, the foreign viral capsid protein used in pseudotyped recombinant lentiviral particles is Vesicular stomatitis virus glycoprotein (VSV-GP). VSV-GP interacts with a ubiquitous cell receptor, providing broad tissue tropism to pseudotyped recombinant lentiviral particles. In addition, VSV-GP is thought to provide higher stability to pseudotyped recombinant lentiviral particles. In other embodiments, the foreign viral capsid proteins are derived from, including without limitation. Chandipura virus. Rabies virus. Mokola virus, Lymphocytic choriomeningitis virus (LCMV). Ross River virus (RRV), Sindbis virus, Semliki Forest virus (SFV), Venezuelan equine encephalitis virus. Ebola virus Reston, Ebola virus Zaire, Marburg virus, Lassa virus, Avian leukosis virus (ALV), Jaagsiekte sheep retrovirus (JSRV). Moloney Murine leukemia virus (MLV). Gibbon ape leukemia virus (GALV). Feline endogenous retrovirus (RD114). Human T-lymphotropic virus 1 (HTLV-1), Human foamy virus, Maedi- visna virus (MW), SARS-CoV, Sendai virus, Respiratory syncytia virus (RSV), Human parainfluenza virus type 3, Hepatitis C virus (HCV), Influenza virus. Fowl plague virus (FPV), or Autographa californica multiple nucleopolyhedro virus (AcMNPV).

[0120] In some embodiments, the recombinant lentiviral vector is derived from a lentivirus pseudotyped with vesicular stomatitis virus (VSV), lymphocytic choriomeningitis virus (LCMV), Ross river virus (RRV), Ebola virus. Marburg virus, Mokala virus, Rabies virus, RD114, or variants therein. Examples of vector and capsid protein combinations used in pseudotyped Lentivirus particles can be found, for example, in Cronin. J. et al. (2005). Curr. Gene Ther. 5(4):387-398. Different pseudotyped recombinant lentiviral particles can be used to optimize transduction of particular target cells or to target specific cell types within a particular target tissue (e.g., a diseased tissue). For example, tissues targeted by specific pseudotyped recombinant lentiviral particles, include without limitation, liver (e.g., pseudotyped with a VSV- G, LCMV, RRV, or SeV F protein), lung (e.g., pseudotyped with an Ebola, Marburg, SeV F and HN, or JSRV protein), pancreatic islet cells (e.g., pseudotyped with an LCMV protein), centralnervous system (e.g., pseudotyped with a VSV-G. LCMV. Rabies, or Mokola protein), retina (e.g., pseudotyped with a VSV-G or Mokola protein), monocytes or muscle (e.g., pseudotyped with a Mokola or Ebola protein), hematopoietic system (e.g., pseudotyped with an RD114 or GALV protein), or cancer cells (e.g., pseudotyped with a GALV or LCMV protein). For further description, see Cronin, J. et al. (2005). Curr. Gene Ther. 5(4):387-398 and Kay, M. et al. (2001) Nat. Med. 7(l):33-40. In some embodiments, the recombinant lentiviral particle comprises a capsid pseudotyped with vesicular stomatitis virus (VSV), lymphocytic choriomeningitis virus (LCMV), Ross river virus (RRV), Ebola virus, Marburg virus, Mokala virus, Rabies virus, RD114 or variants thereof.

[0121] A viral vector can be a lentiviral vector (e.g., human immunodeficiency virus 1 (HIV- 1), human immunodeficiency virus 2 (HIV-2), visna-maedi virus (VMV) virus, caprine arthritisencephalitis virus (CAEV), equine infectious anemia virus (EIAV), feline immunodeficiency virus (FIV), bovine immune deficiency virus (BIV), simian immunodeficiency virus (SIV), derivatives thereof, or any combination thereof). A viral vector can be a recombinant lentiviral vector. The recombinant lentiviral vector can be derived from a lentivirus pseudotyped with vesicular stomatitis virus (VSV), lymphocytic choriomeningitis virus (LCMV), Ross river virus (RRV), Ebola virus, Marburg virus, Mokala virus, Rabies virus, RD114, or variants therein. The viral vector composition can comprise: one or more of a left (5') retroviral LTR, a Psi ( ) packaging signal, a central polypurine tract / DNA flap (cPPT / FLAP), a retroviral export element, and a right (3') retroviral LTR. The promoter of the 5' LTR can be replaced with a heterologous promoter. The 5' LTR or 3' LTR can be a lentivirus LTR. The 3' LTR can comprise one or more modifications and / or deletions. The 3' LTR can be a self-inactivating (SIN) LTR.

[0122] In some embodiments, the viral vector is a recombinant adenoviral vector. In some embodiments, the viral particle is an adenoviral particle. In some embodiments, the adenoviral particle is a recombinant adenoviral particle. In some embodiments, the adenoviral particle lacks or contains a defective copy of one or more El genes, which renders the adenovirus replicationdefective. Adenoviruses include a linear, double-stranded DNA genome within a large (~950 A), non-enveloped icosahedral capsid. Adenoviruses have a large genome that can incorporate more than 30 kb of heterologous sequence (e.g., in place of the El and / or E3 region), making them uniquely suited for use with larger heterologous genes. They are also known to infect dividing and non-dividing cells and do not naturally integrate into the host genome (although hybrid variants may possess this ability). In some embodiments, the adenoviral vector may be a first generation adenoviral vector with a heterologous sequence in place of EL In some embodiments, the adenoviral vector may be a second generation adenoviral vector with additional mutations ordeletions in E2A. E2B, and / or E4. In some embodiments, the adenoviral vector may be a third generation or gutted adenoviral vector that lacks all viral coding genes, retaining only the ITRs and packaging signal and requiring a helper adenovirus in trans for replication, and packaging. Adenoviral particles have been investigated for use as vectors for transient transfection of mammalian cells as well as gene therapy vectors. For further description, see. e.g., Danthinne. X, and Imperiale, M. J. (2000) Gene Ther. 7: 1707-14 and Tatsis. N, and Ertl. H. C. (2004) Mol. Ther. 10:616-29.

[0123] In some embodiments, the viral vector is an rHSV vector. In some embodiments, the viral particle is a herpes simplex virus (HSV) particle. In some embodiments, the HSV particle is a rHSV particle. HSV is an enveloped, double-stranded DNA virus with a genome of approximately 152 kb. Advantageously, approximately half of its genes are nonessential and may be deleted to accommodate heterologous sequence. HSV particles infect non-dividing cells. In addition, they naturally establish latency in neurons, travel by retrograde transport, and can be transferred across synapses, making them advantageous for transfection of neurons and / or gene therapy approaches involving the nervous system. In some embodiments, the HSV particle may be replication-defective or replication-competent (e.g., competent for a single replication cycle through inactivation of one or more late genes). For further description, see Manservigi, R. et al. (2010) Open Virol. J. 4: 123-56.

[0124] The AAV vector can comprise single-stranded AAV (ssAAV) vector or a self- complementary AAV (scAAV) vector. The AAV vector can comprise AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, derivatives thereof, or any combination thereof. The AAV vector can comprise an AAV9 variant engineered for systemic delivery (e.g., AAVPHP.B, AAV-PHP.eB, or AAV-PHP.S). The AAV vector can be or can comprise an AAV selected from the group consisting of AAV9, AAV9 K449R (or K449R AAV9), AAV1, AAVrhlO, AAV-DJ, AAV-DJ8, AAV5, AAVPHP.B (PHP.B), AAVPHP. A (PHP. A), AAVG2B-26, AAVG2B-13, AAVTH1.1-32, AAVTH1.1- 35, AAVPHP.B2 (PHP.B2), AAVPHP.B 3 (PHP.B 3), AAVPHP.N / PHP.B-DGT, AAVPHP.B-EST, AAVPHP. B-GGT, AAVPHP. B-ATP, AAVPHP. B-ATT-T, AAVPHP.B- DGT-T, AAVPHP. B-GGT-T, AAVPHP. B-SGS, AAVPHP.B-AQP, AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B- SNP, AAVPHP.B-QGT, AAVPHP. B-NQT, AAVPHP.B- EGS, AAVPHP.B-SGN, AAVPHP.B- EGT, AAVPHP. B-DST, AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP, AAVPHP.B- SQP, AAVPHP.B-QLP, AAVPHP.B- TMP, AAVPHP.B-TTP, AAVPHP.S / G2A12, AAVG2 Al 5 / G2A3 (G2A3), AAVG2B4 (G2B4), AAVG2B5 (G2B5), PHP.S, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7,AAV7.2, AAV8, AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11, AAV 12, AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42- lb, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42- aa, AAV43-1, AAV43-12, AAV43-20, AAV43-21, AAV43-23, AAV43-25, AAV43-5, AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7, AAVl-7 / rh.48, AAVl-8 / rh.49, AAV2-15 / rh.62, AAV2-3 / rh.61, AAV2-4 / rh.5O, AAV2-5 / rh.51, AAV3. l / hu.6, AAV3. l / hu.9, AAV3-9 / rh.52, AAV3-1 l / rh.53, AAV4- 8 / rl 1.64, AAV4-9 / rh.54, AAV4-19 / rh.55, AAV5-3 / rh.57, AAV5-22 / rh.58, AAV7.3 / hu.7, AAV16.8 / hu.lO, AAV16.12 / hu.l 1, AAV29.3 / bb.l, AAV29.5 / bb.2, AAV106. l / hu.37, AAV1 14.3 / hu.4O, AAV127.2 / hu.41, AAV127.5 / hu.42, AAV128.3 / hu.44, AAV130.4 / hu.48, AAV145. l / hu.53, AAV145.5 / hu.54, AAV145.6 / hu.55, AAV161. 10 / hu.60, AAV161.6 / hu.61, AAV33. 12 / hu.l7, AAV33.4 / hu.l5, AAV33.8 / hu.l6, AAV52 / hu.l9, AAV52.1 / hu.2O, AAV58.2 / hu.25, AAVA3.3, AAV A3.4, AAV A3.5, AAV A3.7, AAVC1, AAVC2, AAVC5, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70, AAVpi. 1, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu. 12, AAVH6, AAVH-l / hu.l, AAVH-5 / hu.3, AAVLG-10 / rh.40, AAVLG-4 / rh.38, AAVLG-9 / hu.39, AAVN721-8 / rh.43, AAVCh.5, AAVCh.5Rl, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5Rl, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu. 1, AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.lO, AAVhu. 1 1, AAVhu. 13, AAVhu.15, AAVhu.16, AAVhu. 1 7, AAVhu.l 8, AAVhu.20, AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44Rl, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48Rl, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58,AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu. 1 4 / 9, AAVhu.t 19, AAVrh.2, AAVrh.2R, AAVrh.8, AAVrh.8R, AAVrh. 10, AAVrh.12, AAVrh. 13,AAVrh.13R, AAVrh. 14, AAVrh.17, AAVrh. 18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.3 1, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49, AAVrh.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64Rl, AAVrh.64R2, AAVrh.67, AAVrh.73, AAVrh.74, AAVrh8R, AAVrh8R A586R mutant,AAVrh8R R533 A mutant, AAAV, BAAV, caprine AAV, bovine AAV, AAVhEl.l, AAVhErl.5, AAVhERl. 14, AAVhErl.8, AAVhErl. 16, AAVhErl .18, AAVhErl.35, AAVhErl.7, AAVhErl.36, AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.3 1, AAVhEr2.36, AAVhER1.23, AAVhEr3.1, AAV2.5T , AAV-PAEC, AAV-LK01, AAV-LK02, AAV- LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV-LK09, AAV- LK10, AAV-LK11, AAV-LK12, AAV-LK13, AAV-LK14, AAV-LK15, AAV-LK16, AAV-LK17, AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV- PAEC8, AAV-PAEC11, AAV-PAEC12, AAV-2-pre-miRNA-101 , AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2 , AAV Shuffle 100-1 , AAV Shuffle 100-3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM 10-1, AAV SM 10-8 , AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrh.48, AAVhu.19, AAVhu. 1 1, AAVhu.53, AAV4- 8 / rh.64, AAVLG-9 / hu.39, AAV54.5 / hu.23, AAV54.2 / hu.22, AAV54.7 / hu.24, AAV54.1 / hu.21, AAV54.4R / hu.27, AAV46.2 / hu.28, AAV46.6 / hu.29, AAV128. l / hu.43, true type AAV (ttAAV), EGRENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7. 10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-El, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV CHt-6. 1, AAV CHt-6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-Pl, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-Bl, AAV CKd-B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV CKd-B8, AAV CKd-Hl, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAV CKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-Fl, AAV CLg-F2, AAV CLg-F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1, AAV CLvl-1, AAV Clvl-10, AAV CLvl-2, AAV CLv-12, AAV CLvl-3, AAV CLv-1 3, AAV CLvl-4, AAV Clvl-7, AAV Clvl-8, AAV Clvl-9, AAV CLv- 2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-Dl, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-El, AAV CLv-Kl, AAV CLv-K3, AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv- L6, AAV CLv-Ml, AAV CLv-Ml 1, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv- M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-Rl, AAV CLv-R2, AAV CLv-R3, AAV CLv- R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV CSp-1, AAV CSp-10, AAV CSp-1 1, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8. 10, AAV CSp-8.2, AAV CSp-8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1, AAVF11 / HSC11, AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5, AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, AAVF9 / HSC9, variants thereof, a hybrid or chimera of any of the foregoing AAV serotypes, or any combination thereof.AAV compositions and retinal delivery

[0125] In some embodiments, the AAV or other viral vector disclosed herein is engineered for enhanced transduction to the retina. Methods and compositions for enhanced retinal delivery are also described in PCT publication W02012 / 145601, the contents of which is incorporated by reference herein in its entirety.

[0126] The AAV vector can comprise an AAV variant engineered for reduced binding to heparin sulfate proteoglycans. The AAV vector can comprise an AAV variant engineered for enhanced transduction across the internal limiting membrane (ILM) of the retina. The AAV vector can comprise an AAV variant engineered for enhanced transduction of the retinal cells. In some embodiments, the retinal cells comprise amacrine cells, bipolar cells, rods, cones, cells of the retinal pigment epithelium, or any combination thereof. The AAV vector can comprise a capsid protein comprising an insertion of a targeting peptide after amino acid 587 (AA587) of the AAV capsid protein or a functional equivalent of AA587. In some embodiments, the targeting peptide insertion results in enhanced transduction of retinal cells as compared to an AAV comprising a capsid protein that does not comprise the insertion. The targeting peptide can be about seven amino acids in length. The composition can comprise a recombinant AAV (rAAV).Pharmaceutical Compositions

[0127] Also disclosed herein are pharmaceutical compositions comprising one or more of the viral vector compositions disclosed herein and one or more pharmaceutically acceptable carriers. The compositions can also comprise additional ingredients such as diluents, stabilizers, excipients, and adjuvants. As used herein, “pharmaceutically acceptable” carriers, excipients, diluents, adjuvants, or stabilizers are nontoxic to the cell or subject being exposed thereto (preferably inert) at the dosages and concentrations employed or that have an acceptable level of toxicity as determined by the skilled practitioners. The carriers, diluents and adjuvants can include buffers such as phosphate, citrate, or other organic acids: antioxidants such as ascorbic acid; low molecular weight polypeptides (e.g., less than about 10 residues); proteins such asserum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, di saccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as Tween™, Pluronics™ or polyethylene glycol (PEG). In some embodiments, the physiologically acceptable carrier is an aqueous pH buffered solution.

[0128] Titers of the viral vectors provided herein to be administered will vary depending, for example, on the particular viral vector(s), the mode of administration, the subject, and the cell type(s) being targeted, and can be determined by methods standard in the art. As will be readily apparent to one skilled in the art, the useful in vivo dosage of the viral vector(s) to be administered and the particular mode of administration will vary depending upon the age, weight, and animal species treated, the particular viral vector(s) that is used, and the specific use for which the viral vector(s) is employed. The determination of effective dosage levels, that is the dosage levels necessary to achieve the desired result, can be accomplished by one skilled in the art using routine pharmacological methods.

[0129] In some embodiments, pharmaceutical compositions in accordance with the present disclosure are administered at dosage levels sufficient to deliver from about 0.0001 mg / kg to about 100 mg / kg, from about 0.001 mg / kg to about 0.05 mg / kg, from about 0.005 mg / kg to about 0.05 mg / kg, from about 0.001 mg / kg to about 0.005 mg / kg, from about 0.05 mg / kg to about 0.5 mg / kg, from about 0.01 mg / kg to about 50 mg / kg, from about 0.1 mg / kg to about 40 mg / kg, from about 0.5 mg / kg to about 30 mg / kg, from about 0.01 mg / kg to about 10 mg / kg, from about 0.1 mg / kg to about 10 mg / kg, or from about 1 mg / kg to about 25 mg / kg, of subject body weight per day, one or more times a day, to obtain the effect. It will be understood that the above dosing concentrations may be converted to vg or viral genomes per kg or into total viral genomes administered by one of skill in the art.

[0130] In some embodiments, a dose of the pharmaceutical composition comprises a concentration of infectious particles of at least or about 107, 108, 109, 1010, 1011, 1012, 1013, 1014, 1015, 1016, or IO17In some cases the concentration of infectious particles is 2* 107, 2* 108, 2x l09, 2x lO10, 2* 1011, 2* 1012, 2* 1013, 2* 1014, 2* 1015, 2* 1016, 2* 1017, or a range between any two of these values. In some cases the concentration of the infectious particles is 3* 107, 3* 108, 3* 109, 3x l010, 3x 1011, 3x l012, 3x l013, 3x l014, 3x 1015, 3x l016, 3x 1017, or a range between any two of these values. In some cases the concentration of the infectious particles is 4x l07, 4x l08, 4x l09, 4x iQ10, 4x io1, 4x l012, 4x l013, 4x 1014, 4x l015, 4x l016, 4x l017, or a range between any two ofthese values. In some cases the concentration of the infectious particles is 5* 107, 5* 108, 5* 109, 5x lO10, 5* 1011, 5* 1012, 5* 1013, 5* 1014, 5* 1015, 5* 1016, 5* 1017, or a range between any two of these values. In some cases the concentration of the infectious particles is 6* 107, 6* 108, 6* 109, 6x lO10, 6x 1011, 6x l012, 6x l013, 6x l014, 6x 1015, 6x l016, 6x 1017, or a range between any two of these values. In some cases the concentration of the infectious particles is 7x l07, 7x l08, 7x l09, 7x lO10, 7x lOn, 7x l012, 7x l013, 7x lO147x l015, 7x l06, 7x l017, or a range between any two of these values. In some cases the concentration of the infectious particles is 8x l07, 8x l08, 8x l09, 8x lO10, 8x 1011, 8x l012, 8x l013, 8x lO148x 1015, 8x l016, 8x 1017, or a range between any two of these values. In some cases the concentration of the infectious particles is 9x l07, 9x l08, 9x l09, 9x lO10, 9x 1011, 9x l012, 9x l013, 9x lO149x 1015, 9x l016, 9x 1017, or a range between any two of these values.Methods of Treating Retinal Diseases or Disorders

[0131] Photoreceptors are the first neurons in the retina to receive and process visual information, converting visible electromagnetic radiation into hyperpolarized responses through phototransduction. The overwhelming majority of inherited retinal diseases result in the loss of these cells, either directly, such as in dominant mutations that affect rhodopsin protein folding, or indirectly, such as in recessive mutations that affect retinal recycling pathways in the retinal pigment epithelium (RPE).

[0132] The nucleic acids described herein can comprise sequence encoding a gene product (e.g., a therapeutic gene product). In some embodiments, the gene product is an interfering RNA. In some embodiments, the gene product is an aptamer. In some embodiments, the gene product is a polypeptide. In some embodiments, the gene product is a site-specific nuclease that provide for site-specific knock-down of gene function.

[0133] Interfering RNA (RNAi), can include, but is not limited to RNAi that decrease the level of an apoptotic or angiogenic factor in a cell. For example, an RNAi can be an shRNA or siRNA that reduces the level of a gene product that induces or promotes apoptosis in a cell. Genes whose gene products induce or promote apoptosis are referred to herein as "pro-apoptotic genes" and the products of those genes (mRNA; protein) are referred to as "pro-apoptotic gene products." Pro-apoptotic gene products include, e.g., Bax, Bid, Bak, and Bad gene products. See, e.g., U.S. Patent No. 7,846,730. In some embodiments, interfering RNAs can be against an angiogenic product, for example, VEGF (e.g., Cand5), or VEGFR2. In some embodiments, the gene product comprises aptamers. Exemplary aptamers of interest include an aptamer against vascular endothelial growth factor (VEGF). In some embodiments the gene product is apolypeptide (e.g., a protein or fragment thereof). In some embodiments, the polypeptide enhances function of a retinal cell, e.g., the function of a rod or cone photoreceptor cell, a retinal ganglion cell, a Muller cell, a bipolar cell, an amacrine cell, a horizontal cell, or a retinal pigmented epithelial cell. Exemplary polypeptides include neuroprotective polypeptides (e.g., GDNF, CNTF, NT4, NGF, and NTN); anti -angiogenic polypeptides (e.g., a soluble vascular endothelial growth factor (VEGF) receptor; a VEGF -binding antibody; a VEGF -binding antibody fragment (e.g., a single chain anti- VEGF antibody); endostatin; tumstatin; angiostatin; a soluble Fit polypeptide; an Fc fusion protein comprising a soluble Fit polypeptide; pigment epithelium- derived factor (PEDF); a soluble Tie-2 receptor; etc.); tissue inhibitor of metalloproteinases-3 (TIMP-3); a light-responsive opsin, e.g., a rhodopsin; anti- apoptotic polypeptides (e.g., Bcl-2, Bcl-Xl); and the like. Suitable polypeptides include, but are not limited to, glial derived neurotrophic factor (GDNF); fibroblast growth factor 2; neurturin (NTN); ciliary neurotrophic factor (CNTF); nerve growth factor (NGF); neurotrophin-4 (NT4); brain derived neurotrophic factor (BDNF); epidermal growth factor; rhodopsin; X-linked inhibitor of apoptosis; and Sonic hedgehog. In some embodiments, the therapeutic gene product comprises a suitable light-responsive opsins include, e.g., a light-responsive opsin as such as channel rhodopsin.. In some embodiments, the polypeptides comprises retinoschisin). In some embodiments, the polypeptide comprises, e.g., retinitis pigmentosa GTPase regulator (RGPR)- interacting protein-1 (see, e.g., GenBank Accession Nos. Q96KN7, Q9EPQ2, and Q9GLM3); peripherin-2 (Prph2) (see, e.g., GenBank Accession No. NP_000313; peripherin; a retinal pigment epithelium- specific protein (RPE65).

[0134] In some embodiments, the polypeptide comprises, CHM (choroidermia (Rab escort protein 1)), a polypeptide that, when defective or missing, causes choroideremia. In some embodiments, the polypeptide comprises Crumbs homolog 1 (CRB1), a polypeptide that, when defective or missing, causes Leber congenital amaurosis and retinitis pigmentosa (see, e.g., den Hollander et al. (1999) Nat. Genet. 23:217; and GenBank Accession No. CAM23328).

[0135] In some embodiments, the polypeptide comprises polypeptide that, when defective or missing, lead to achromotopsia, where such polypeptides include, e.g., cone photoreceptor cGMP-gated channel subunit alpha (CNGA3) (see, e.g., GenBank Accession No. NP_001289); guanine nucleotide binding protein (G protein), alpha transducing activity polypeptide 2 (GNAT2) (ACHM4); and ACHM5; and polypeptides that, when defective or lacking, lead to various forms of color blindness (e.g., L-opsin, M-opsin, and S-opsin).

[0136] In some embodiments, methods and compositions are provided for treatment of macular degeneration. In wet age-regulated macular degeneration, atypical blood vessels appeararound the macula and retina. These vessels can leak proteins or lipids, and they may also cause scarring. Anti -angiogenic therapies can be effective in treating diseases such as AMD. Vascular endothelial growth factor (VEGF) is the main protein that causes new blood vessels to form in the macula in wet AMD. VEGF inhibitors reduce leakage from blood vessels, prevent their growth, decrease swelling of the retina, reduce vision loss, and improve vision in wet AMD.

[0137] Dry AMD is a slow deterioration of the cells of the macula, often over many years, as the retinal cells die off and are not renewed. Without being bound by any particular theory, the complement system is thought to play a role in the progression of dry AMD. Most of the proteins and glycoproteins that constitute the complement system are synthesized by hepatocytes, but significant amounts are also produced by tissue macrophages, blood monocytes, and epithelial cells of the genitourinary system and gastrointestinal tract. The three pathways of activation all generate homologous variants of the protease C3-convertase. The classical complement pathway typically requires antigen-antibody complexes for activation (specific immune response), whereas the alternative pathway can be activated by spontaneous complement component 3 (C3) hydrolysis, foreign material, pathogens, or damaged cells. The mannose-binding lectin pathway can be activated by C3 hydrolysis or antigens without the presence of antibodies (non-specific immune response). In all three pathways, C3-convertase cleaves and activates component C3, creating C3a and C3b, and causes a cascade of further cleavage and activation events. C3b binds to the surface of pathogens, leading to greater internalization by phagocytic cells by opsonization.

[0138] Uveitis is a form of eye inflammation. It affects the middle layer of tissue in the eye wall (uvea). Uveitis warning signs often come on suddenly and get worse quickly. They include eye redness, pain and blurred vision. The condition can affect one or both eyes, and it can affect people of all ages, including children. Causes of uveitis include infection, injury, or an autoimmune or inflammatory disease. In some embodiments the cause is not identified (e.g., is idiopathic). Uveitis can be serious, leading to permanent vision loss.

[0139] Disclosed herein include methods of treating a retinal disease or disorder in a subject. In some embodiments, the method comprises: administering any of the compositions disclosed herein to the subject; and applying thermal energy to a target site of the subject sufficient to increase local temperature of the target site to an activating temperature, thereby inducing the expression of the therapeutic gene product.

[0140] The target site can comprise the retina. The composition can be administered at a dose of about 1 x 109viral genomes to about 1.0 x 1012viral genomes (e.g., 1 x io9, 1 x io10, 1 x 1011, 1 x io12, viral genomes, or a number or range between any two of these values). Thecomposition can be administered at a dose of more than about 1.0 * 1012viral genomes. The composition can be administered by intravitreal route, subretinal route, or suprachoroidal route.

[0141] The method can comprise a single administration of the composition to the subject. The method can comprise administering the composition two or more times to the subject. Each of the two or more administrations can be 1 month, 2 months, 6 months, 1 year, or more, apart. The period of time between the administering the composition and applying thermal energy can be about 1 week, about 6 days, about 5 days, about 4 days, about 3 days, about 48 hours, about 44 hours, about 40 hours, about 35 hours, about 30 hours, about 25 hours, 20 hours, 15 hours, 10 hours, about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, about 10 minutes, or about 5 minutes. The method can comprise two or more applications of thermal energy (e.g., daily, weekly, or monthly; or in response to flare in symptoms). The period of time between administering the composition and applying thermal energy can be the period of time between the administering the composition and the first application of thermal energy.

[0142] In some embodiments, the method comprises: applying thermal energy to a target site of the subject sufficient to increase the local temperature of the target site to an activating temperature, thereby inducing the expression of the therapeutic gene product. The activating temperature can be greater than 37°C. The activating temperature can comprise a temperature of at least 37.5° C to at most 70°C. The activating temperature can be about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, about 46.0°C, about 46.5°C, about 47.0°C, about 47.5°C, about 48.0°C, about 48.5 °C, about 49.0°C, about 49.5°C, about 50.0°C, about 50.5°C, about 51.0°C, about 51.5°C, about 52.0°C, about 52.5°C, about 53.0°C, about 53.5°C, about 54.0°C, about 54.5°C, about 55.0°C, about 55.5°C, about 56.0°C, about 56.5°C, about 57.0°C, about 57.5°C, about 58.0°C, about 58.5°C, about 59.0°C, about 59.5°C, about 60.0°C, about 60.5°C, about 61.0°C, about 61.5°C, about 62.0°C, about 62.5°C, about 63.0°C, about 63.5°C, about 64.0°C, about 64.5°C, about 65.0°C, about 65.5°C, about 66.0°C, about 66.5°C, about 67.0°C, about 67.3°C, about 67.5°C, about 68.0°C, about 68.5°C, about 69.0°C, about 69.5°C, or about 70.0°C. The thermal energy can be applied to the target site for a duration of time comprising about 1 second, about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, or about 1 minute.

[0143] The activating temperature can be about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, or about 46.0°C. The thermal energy can be applied to the target site for a duration of time comprising about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes.

[0144] The activating temperature can comprise a temperature of about 42°C to about 43°C. The activating temperature can be about 42.0°C, about 42.1°C, about 42.2°C, about 42.3°C, about 42.4°C, about 42.5°C, about 42.6°C, about 42.7°C, about 42.8°C, about 42.9°C, or about 43.0°C.

[0145] The thermal energy can be applied to the target site for a duration of time comprising about 1 second to about 20 minutes. The thermal energy can be applied to the target site for a duration of time comprising about 1 second, about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes.

[0146] The temperature at the target site can be elevated to about 42°C to about 43 °C after the applying. The temperature at the target site can be elevated to about 42°C to about 43 °C within at most 30 minutes from the applying. The temperature at the target site can be elevated to about 42°C to about 43 °C within 10 minutes or less after the applying. The temperature at the target site can be elevated to about 42°C to about 43 °C within about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, or about 10 minutes after the applying. The temperature at the target site can be maintained at about 42°C to about 43°C for at least about 15 minutes to at least about 120 minutes.

[0147] The temperature at the target site can be maintained at 42°C to 43°C for at least 15 minutes, at least 20 minutes, at least 25 minutes, at least 30 minutes, at least 35 minutes, at least 40 minutes, at least 45 minutes, at least 50 minutes, at least 55 minutes, at least 60 minutes, at least 90 minutes, or at least 120 minutes. The temperature at the target site can be maintained at 42°C to 43 °C for at least 20 minutes. The temperature at the target site can be maintained atabout 42°C to about 43 °C for about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes. Applying thermal energy to the target site of the subject can comprise the application of one or more of ciliary body hyperthermia, magnetic hyperthermia, microwave hyperthermia, inductive diathermy, contact hyperthermia, ultrasonic diathermy, focused ultrasound, and deep hyperthermia. Applying thermal energy to the target site of the subject can comprise use of a laser. The laser can be a 577 nm or a 532 nm laser. The laser can be set at a power level of about 25% to about 40%. The laser can be set at a power level of about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, or about 40%. The laser can be set at a power level of about 30%. The laser can be applied to the target site for a pulse of about 1 second to about 15 seconds. The laser can be applied to the target site for a pulse of about 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 11 seconds, 12 seconds, 13 seconds, 14 seconds, or 15 seconds.

[0148] In some embodiments, upon applying thermal energy to the target site, the transcription of the therapeutic gene, the transactivator gene, the recombinase gene, or any combination thereof, increases by at least 1.1-fold (e.g., 1.1-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or a number or a range between any of these values). In some embodiments, upon applying thermal energy to the target site, the levels of the therapeutic gene product, the transactivator gene product, the recombinase gene product, or any combination thereof, increases by at least 1.1-fold (e.g., 1.1-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6- fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or a number or a range between any of these values), wherein the gene product comprises RNA transcribed from the gene. The gene product can comprise an mRNA, a protein translated from the mRNA, or both.

[0149] Central retinal thickness (CRT), best-corrected visual acuity (BCVA) or both, can be improved in the subject following the applying of thermal energy. The method can comprise applying thermal energy to the subject two or more times. Each of the two or more applications of thermal energy can be 1 week apart, 2 weeks apart, 3 weeks apart, 4 weeks apart, 5 weeks apart, or more. The time between each of the two or more applications of thermal energy can be the same or different. In some embodiments, each of the two or more applications of thermalenergy can be applied on a schedule, as needed (e.g., in response to flares), or both. In some embodiments, the schedule can be daily, weekly, or monthly. Each of the two or more applications of thermal energy can be applied when: (a) the best-corrected visual acuity (BCVA) of the subject decreases by at least 5 letters, and / or (b) the central retinal thickness CRT increases by at least 100 pM. In some embodiments, the decrease of (a) and the increase of (b) is relative to the BCVA and CRT (i) in a subject that has not been administered the composition; (ii) prior to administering the composition to the subject; (iii) prior to the first application of thermal energy to the subject, or (iv) following a prior application of the thermal energy to the subject.

[0150] The retinal disease can comprise age-related macular degeneration (AMD), non- infectious uveitis, Choroidemia, Leber congenital amaurosis (LCA), Leber hereditary optic neuropathy (LHON), Retinitis pigmentosa (RP), Achromatopsia, or Retinoscisis. The AMD can be wet or dry AMD.EXEMPLARY EMBODIMENTS1. A composition comprising: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a polynucleotide comprising a gene encoding a gene product; wherein the first inducible promoter is capable of inducing transcription of the gene to generate the gene product upon thermal stimulation.2. A composition comprising: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a transactivator gene, and a second promoter operably linked to a second polynucleotide comprising a gene, wherein the first inducible promoter is capable of inducing transcription of the transactivator gene to generate a transactivator transcript in the presence of thermal stimulation, wherein the transactivator transcript is capable of being translated to generate a transactivator; and wherein, in the presence of the transactivator and optionally, a transactivator-binding compound, the second promoter is capable of inducing transcription of a gene to generate a gene product.3. A composition comprising:a viral vector comprising a nucleic acid comprising a first inducible promoter and a second promoter each operably linked to a first polynucleotide comprising a gene and to a second polynucleotide comprising a transactivator gene, wherein the first inducible promoter is capable of inducing transcription of the gene and the transactivator gene to generate a polycistronic transcript upon thermal stimulation, wherein, in the presence of the transactivator and a transactivator-binding compound, the second promoter is capable of inducing transcription of the gene and the transactivator gene to generate a polycistronic transcript, and wherein the polycistronic transcript is capable of being translated to generate a transactivator and a protein.4. The composition of any one of embodiments 2-3, wherein the second promoter comprises one or more copies of a transactivator recognition sequence the transactivator is capable of binding to induce transcription, and wherein the transactivator is incapable of binding the transactivator recognition sequence in the absence of the transactivator-binding compound, optionally the one or more copies of a transactivator recognition sequence comprise one or more copies of a tet operator (TetO).5. The composition of any one of embodiments 2-4, wherein the transactivator comprises reverse tetracycline-controlled transactivator (rtTA).6. The composition of any one of embodiments 2-4, wherein the transactivator comprises tetracycline-controlled transactivator (tTA).7. The composition of any one of embodiments 2-6, wherein the transactivatorbinding compound comprises tetracycline, doxycycline or a derivative thereof.8. The composition of any one of embodiments 3-7, wherein the first polynucleotide and the second polynucleotide are operably linked to a tandem gene expression element, optionally the tandem gene expression element is an internal ribosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof.9. A composition comprising: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a recombinase gene, wherein the first inducible promoter is capable of inducing transcription of the recombinase gene to generate a recombinase transcript upon thermal stimulation,and wherein the recombinase transcript is capable of being translated to generate a recombinase; a second promoter and a second polynucleotide comprising a gene, wherein, in the absence of a recombination event, the second promoter and the second polynucleotide are not operably linked, wherein the recombinase is capable of catalyzing the recombination event, and wherein the second promoter and the second polynucleotide are operably linked after the recombination event such that the second promoter is capable of inducing transcription of the gene to generate a gene product.10. The composition of embodiment 9, wherein the recombination event comprises removal of a sequence flanked by recombinase target sites or an inversion of a sequence flanked by recombinase target sites; optionally the second polynucleotide is flanked by recombinase target sites.11. The composition of any one of embodiments 9-10, wherein, prior to the recombination event, the sequence of the gene is inverted relative to the promoter.12. The composition of any one of embodiments 9-11, comprising at least one stop cassette situated between the second promoter and the gene, wherein the stop cassette comprises one or more stop sequences, and wherein the one or more stop cassettes are flanked by recombinase target sites.13. The composition of any one of embodiments 1-12, wherein the gene product is an RNA transcript and / or protein, optionally the transcript is an mRNA capable of being translated to generate the protein.14. The composition of any one of embodiments 12-13, wherein the at least one stop cassette is configured to prevent transcription of the gene and / or translation of the transcript.15. The composition of any one of embodiments 12-14, wherein the one or more stop sequences comprise a polyadenylation signal, a stop codon, a frame-shifting mutation, or any combination thereof.16. The composition of any one of embodiments 2-15, wherein the second promoter comprises a ubiquitous or tissue-specific promoter.17. The composition of any one of embodiments 2-16, wherein the second promoter comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focusforming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Roussarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl l promoters from vaccinia virus, an elongation factor 1-alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3- phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P- kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CBH promoter, any variant thereof, or any combination thereof.18. The composition of any one of embodiments 9-17, wherein the recombinase is Cre, Dre, Flp, KD, B2, B3, , HK022, HP1, y6, ParA, Tn3, Gin, OC31, Bxbl, R4, derivatives thereof, or any combination thereof; optionally the recombinase is a Flp recombinase and the recombinase target sites are FRT sites or the recombinase is a Cre recombinase and the recombinase target sites are loxP sites.19. The composition of any one of embodiments 1-18, wherein the first inducible promoter comprises a core promoter and at least one heat shock element (HSE).20. The composition of embodiment 19, wherein the core promoter comprises the heat shock protein transcription start site of a gene selected from the group consisting of HSPA1A, HSPH1, HSPB1, HSPA6, and YB.21. The composition of any one of embodiments 19-20, wherein the core promoter comprises the sequence of any one of SEQ ID NOs: 9-12 or a sequence having at least 85% identity to any one of SEQ ID NOs: 9-12.22. The composition of any one of embodiments 19-20, wherein the core promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 9-12.23. The composition of any one of embodiments 19-22, wherein the first inducible promoter comprises seven HSEs.24. The composition of any one of embodiments 19-23, wherein each of the at least one HSE comprises the sequence of 5’ - nGAAnnTTCnnGAAn-3’.25. The composition of any one of embodiments 19-24, wherein the at least one HSE comprises the sequence of any one of SEQ ID NOs: 1-8 or a sequence having at least 85% identity to any one of SEQ ID NOs: 1-8 or a sequence differing by one or two nucleotide mismatches relative to any one of SEQ ID NOs: 1-8.26. The composition of any one of embodiments 19-24, wherein the at least one HSE comprises a sequence selected from the group consisting of SEQ ID NOs: 1-8.27. The composition of any one of embodiments 1-18, wherein the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 13-20 or a sequence having at least 85% identity to any one of SEQ ID NOs: 13-20.28. The composition of any one of embodiments 1-18, wherein the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 13-20.29. The composition of any one of embodiments 1-18, wherein the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 21-34 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-34.30. The composition of any one of embodiments 1-18, wherein the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-34.31. A composition comprising: a viral vector comprising: a nucleic acid comprising a leaky inducible promoter operably linked to a polynucleotide comprising a gene encoding a gene product, wherein the leaky inducible promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the gene product; and wherein the leaky inducible promoter is capable of boosting transcription of the gene, thereby driving a second level of expression of the gene product upon thermal stimulation.32. The composition of embodiment 31, wherein the leaky inducible promoter comprises an HSPB, HSPB’2, HSP A / A, or HSP A / B promoter.33. The composition of any one of embodiments 31-32, wherein the leaky inducible promoter comprises the sequence of any one of SEQ ID NOs: 21-24 and 28-29 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-24 and 28-29.34. The composition of any one of embodiments 31-32, wherein the leaky inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-24 and 28- 29.35. A composition comprising: a viral vector comprising: a nucleic acid comprising a tandem promoter operably linked to a polynucleotide comprising a gene encoding a gene product, wherein the tandem promoter comprises a first inducible promoter and a second constitutive promoter,wherein the second constitutive promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the gene product; and wherein the first inducible promoter is capable of boosting transcription of the gene upon thermal stimulation, thereby driving a second level of expression of the gene product.36. The composition of embodiment 35, wherein the tandem promoter comprises, from 5’ to 3’ : the first inducible promoter then the second constitutive promoter; or wherein the tandem promoter comprises, from 5’ to 3’ : the second constitutive promoter then the first inducible promoter.37. A composition comprising: a viral vector comprising: a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a gene encoding a gene product and a second constitutive promoter operably linked to a second polynucleotide comprising the gene encoding a gene product wherein the second constitutive promoter induces transcription of the gene encoding a gene product from the second polynucleotide in the absence of thermal stimulation, thereby driving a first level of expression of the gene product; and wherein the first inducible promoter is capable of inducing the transcription of the gene from the first polynucleotide upon thermal stimulation, thereby boosting the expression of the gene product to a second level of expression.38. The composition of embodiment 37, wherein the nucleic acid comprises, from 5’ to 3’ : the first inducible promoter, the first polynucleotide, the second constitutive promoter, and the second polynucleotide; or wherein the nucleic acid comprises, from 5’ to 3’ : the second constitutive promoter, the second polynucleotide, the first inducible promoter, and the first polynucleotide.39. The composition of any one of embodiments 31-38, wherein the second level of expression is at least 1.1-fold higher than the first level of expression.40. The composition of any one of embodiments 31-39, wherein, after a first duration of time, the gene product returns to the first level of expression from the second level of expression, wherein the first duration of time is about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks or more.41. A composition comprising: a viral vector comprising:a nucleic acid comprising a bidirectional promoter comprising a first inducible promoter in a first orientation, and a second constitutive promoter in a second orientation, wherein the first inducible promoter is operably linked to a first polynucleotide comprising a first gene encoding a gene product, and wherein the second constitutive promoter is operably linked to a second polynucleotide comprising a second gene, wherein the first and second orientations are opposite of each other; wherein the second constitutive promoter is capable of inducing transcription of the second gene in the absence of thermal stimulation; and wherein the first inducible promoter is capable of inducing transcription of the first gene upon thermal stimulation to generate the gene product.42. The composition of embodiment 41, wherein the second gene encodes a gene product; optionally, the gene product is capable of inhibiting vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6).43. The composition of any one of embodiments 35-41, wherein the second constitutive promoter comprises a ubiquitous or tissue-specific promoter.44. The composition of any one of embodiments 35-42, wherein the second constitutive promoter comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focus-forming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl l promoters from vaccinia virus, an elongation factor 1-alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3 -phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P-kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CBH promoter, or any combination thereof.45. The composition of any one of embodiments 35-44, wherein the first inducible promoter comprises a core promoter and at least one heat shock element (HSE).46. The composition of embodiment 45, wherein the core promoter comprises the heat shock protein transcription start site of a gene selected from the group consisting of HSPA1A, HSPH1, HSPB1, HSPA6, and YB.47. The composition of any one of embodiments 45-46, wherein the core promoter comprises the sequence of any one of SEQ ID NOs: 9-12 or a sequence having at least 85% identity to any one of SEQ ID NOs: 9-12.48. The composition of any one of embodiments 45-46, wherein the core promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 9-12.49. The composition of any one of embodiments 45-48, wherein the first inducible promoter comprises seven HSEs.50. The composition of any one of embodiments 45-49, wherein each of the at least one HSE comprises the sequence of 5’ - nGAAnnTTCnnGAAn-3’.51. The composition of any one of embodiments 45-50, wherein the at least one HSE comprises the sequence of any one of SEQ ID NOs: 1-8 or a sequence having at least 85% identity to any one of SEQ ID NOs: 1-8 or a sequence differing by one or two nucleotide mismatches relative to any one of SEQ ID NOs: 1-8.52. The composition of any one of embodiments 45-50, wherein the at least one HSE comprises a sequence selected from the group consisting of SEQ ID NOs: 1-8.53. The composition of any one of embodiments 35-44, wherein the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 13-20 or a sequence having at least 85% identity to any one of SEQ ID NOs: 13-20.54. The composition of any one of embodiments 35-44, wherein the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 13-20.55. The composition of any one of embodiments 35-44, wherein the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 21-34 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-34.56. The composition of any one of embodiments 35-44, wherein the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-34.57. The composition of any one of embodiments 1-56, wherein any two of the first inducible promoter, the second promoter, the leaky inducible promoter, the tandem promoter, the second constitutive promoter, and the bidirectional promoter are in opposite orientations or the same orientation relative to each other.58. The composition of any one of embodiments 1-57, wherein the gene product comprises an siRNA, an shRNA, a miRNA, or a protein.59. The composition of any one of embodiments 1-58, wherein the gene product is capable of inhibiting vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6).60. The composition of embodiment 59, wherein the gene product comprises an antibody or fragment thereof capable of binding to vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6).61. The composition of any one of embodiments 1-60, wherein the thermal stimulation comprises heating to an activating temperature, wherein the activating temperature is greater than 37°C.62. The composition of embodiment 61, wherein the activating temperature comprises a temperature of at least 37°C to at most 70°C.63. The composition of embodiment 62, wherein the activating temperature is about 37.0°C, 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, about 46.0°C, about 46.5°C, about 47.0°C, about 47.5°C, about 48.0°C, about 48.5 °C, about 49.0°C, about 49.5°C, about 50.0°C, about 50.5°C, about 51.0°C, about 51.5°C, about 52.0°C, about 52.5°C, about 53.0°C, about 53.5°C, about 54.0°C, about 54.5°C, about 55.0°C, about 55.5°C, about 56.0°C, about 56.5°C, about 57.0°C, about 57.5°C, about 58.0°C, about 58.5°C, about 59.0°C, about 59.5°C, about 60.0°C, about 60.5°C, about 61.0°C, about 61.5°C, about 62.0°C, about 62.5°C, about 63.0°C, about 63.5°C, about 64.0°C, about 64.5°C, about 65.0°C, about 65.5°C, about 66.0°C, about 66.5°C, about 67.0°C, about 67.3°C, about 67.5°C, about 68.0°C, about 68.5°C, about 69.0°C, about 69.5°C, or about 70.0°C.64. The composition of any one of embodiments 61-63, wherein the activating temperature is about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, or about 46.0°C.65. The composition of any one of embodiments 61-64, wherein the activating temperature comprises a temperature of about 42°C to about 43°C.66. The composition of embodiment 65, wherein the activating temperature is about 42.0°C, about 42.1°C, about 42.2°C, about 42.3°C, about 42.4°C, about 42.5°C, about 42.6°C, about 42.7°C, about 42.8°C, about 42.9°C, or about 43.0°C.67. The composition of any one of embodiments 1-66, wherein the viral vector is or comprises an AAV vector, a lentivirus vector, a retrovirus vector, an adenovirus vector, a herpesvirus vector, a herpes simplex virus vector, a cytomegalovirus vector, a vaccinia virus vector, a MVA vector, a baculovirus vector, a vesicular stomatitis virus vector, a human papillomavirus vector, an avipox virus vector, a Sindbis virus vector, a VEE vector, a Measlesvirus vector, an influenza virus vector, a hepatitis B virus vector, an integration-deficient lentivirus (IDLV) vector derivatives thereof, or any combination thereof.68. The composition of embodiment 67, wherein the viral vector is capable of integrating into a mammalian cell genome, and wherein the gene product is capable of being expressed upon integration.69. The composition of any one of embodiments 67-68, wherein the viral vector is a lentiviral vector, optionally human immunodeficiency virus 1 (HIV-1), human immunodeficiency virus 2 (HIV-2), visna-maedi virus (VMV) virus, caprine arthritis-encephalitis virus (CAEV), equine infectious anemia virus (EIAV), feline immunodeficiency virus (FIV), bovine immune deficiency virus (BIV), simian immunodeficiency virus (SIV), derivatives thereof, or any combination thereof.70. The composition of any one of embodiments 67-69, wherein the viral vector is a recombinant lentiviral vector, optionally the recombinant lentiviral vector is derived from a lentivirus pseudotyped with vesicular stomatitis virus (VSV), lymphocytic choriomeningitis virus (LCMV), Ross river virus (RRV), Ebola virus, Marburg virus, Mokala virus, Rabies virus, RD114, or variants thereof.71. The composition of any one of embodiments 67-70, comprising one or more of a left (5') retroviral LTR, a Psi ( ) packaging signal, a central polypurine tract / DNA flap (cPPT / FLAP), a retroviral export element, and a right (3') retroviral LTR.72. The composition of embodiment 71, wherein the 5' LTR or 3' LTR is a lentivirus LTR.73. The composition of any one of embodiments 71-72, wherein the 3' LTR comprises one or more modifications and / or deletions.74. The composition of any one of embodiments 71-73, wherein the 3' LTR is a selfinactivating (SIN) LTR.75. The composition of any one of embodiments 1-74, wherein the viral vector comprises an AAV vector.76. The composition of embodiment 75, wherein the AAV vector comprises a singlestranded AAV (ssAAV) vector or a self-complementary AAV (scAAV) vector.77. The composition of any one of embodiments 75-76, wherein the AAV vector comprises AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, derivatives thereof, or any combination thereof.78. The composition of any one of embodiments 75-77, wherein the AAV vector comprises an AAV9 variant engineered for systemic delivery, optionally AAV-PHP.B, AAV-PHP.eB, or AAV-PHP.S.79. The composition of any one of embodiments 75-78, wherein the AAV vector is or comprises an AAV selected from the group consisting of AAV9, AAV9 K449R (or K449R AAV9), AAV1, AAVrhlO, AAV-DJ, AAV-DJ8, AAV5, AAVPHP.B (PHP.B), AAVPHP.A (PHP. A), AAVG2B-26, AAVG2B-13, AAVTH1.1-32, AAVTH1.1-35, AAVPHP.B2 (PHP.B2), AAVPHP.B 3 (PHP.B3), AAVPHP.N / PHP.B-DGT, AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP, AAVPHP.B-ATT-T, AAVPHP.B-DGT-T, AAVPHP.B-GGT-T, AAVPHP.B- SGS, AAVPHP.B-AQP, AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B-SNP, AAVPHP.B-QGT, AAVPHP.B-NQT, AAVPHP.B- EGS, AAVPHP.B-SGN, AAVPHP.B-EGT, AAVPHP.B-DST, AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP, AAVPHP.B-SQP, AAVPHP.B-QLP, AAVPHP.B- TMP, AAVPHP.B-TTP, AAVPHP.S / G2A12, AAVG2 Al 5 / G2A3 (G2A3), AAVG2B4 (G2B4), AAVG2B5 (G2B5), PHP.S, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11, AAV 12, AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-lb, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa, AAV43-1, AAV43-12, AAV43-20, AAV43-21, AAV43-23, AAV43-25, AAV43-5, AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7, AAVl-7 / rh.48, AAVl-8 / rh.49, AAV2-15 / rh.62, AAV2-3 / rh.61, AAV2-4 / rh.5O, AAV2-5 / rh.51, AAV3. l / hu.6, AAV3. l / hu.9, AAV3-9 / rh.52, AAV3-1 l / rh.53, AAV4-8 / rl 1.64, AAV4-9 / rh.54, AAV4-19 / rh.55, AAV5-3 / rh.57, AAV5-22 / rh.58, AAV7.3 / hu.7, AAV16.8 / hu.lO, AAV16.12 / hu.ll, AAV29.3 / bb.l, AAV29.5 / bb.2, AAV106. l / hu.37, AAV1 14.3 / hu.4O, AAV127.2 / hu.41, AAV127.5 / hu.42, AAV128.3 / hu.44, AAV130.4 / hu.48, AAV145. l / hu.53, AAV145.5 / hu.54, AAV145.6 / hu.55, AAV161. 10 / hu.60, AAV161.6 / hu.61, AAV33. 12 / hu.l7, AAV33.4 / hu.l5, AAV33.8 / hu.l6, AAV52 / hu.l9, AAV52.1 / hu.2O, AAV58.2 / hu.25, AAVA3.3, AAVA3.4, AAVA3.5, AAVA3.7, AAVC1, AAVC2, AAVC5, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70, AAVpi.l, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu. 12, AAVH6, AAVH-l / hu.l, AAVH-5 / hu.3, AAVLG-10 / rh.40, AAVLG-4 / rh.38, AAVLG-9 / hu.39, AAVN721- 8 / rh.43, AAVCh.5, AAVCh.5Rl, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5Rl, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.l, AAVhu.2, AAVhu.3, AAVhu.4,AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.10, AAVhu.ll, AAVhu.13, AAVhu.15, AAVhu.16, AAVhu. 1 7, AAVhu.18, AAVhu.20, AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44Rl, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48Rl, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.l 4 / 9, AAVhu.t 19, AAVrh.2, AAVrh.2R, AAVrh.8, AAVrh.8R, AAVrh. lO, AAVrh.12, AAVrh.13, AAVrh.BR, AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49, AAVrh.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64Rl, AAVrh.64R2, AAVrh.67, AAVrh.73, AAVrh.74, AAVrh8R, AAVrh8R A586R mutant, AAVrh8R R533 A mutant, AAAV, BAAV, caprine AAV, bovine AAV, AAVhEl.l, AAVhErl.5, AAVhER1.14, AAVhErl.8, AAVhErl. 16, AAVhErl.18, AAVhErl.35, AAVhErl.7, AAVhErl.36, AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.3 1, AAVhEr2.36, AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01, AAV-LK02, AAV- LK03, AAV-LK04, AAV- LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV-LK09, AAV- LK10, AAV-LK11, AAV- LK12, AAV-LK13, AAV-LK14, AAV-LK15, AAV-LK16, AAV-LK17, AAV-LK18, AAV- LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV- PAEC11, AAV-PAEC 12, AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV Shuffle 100-1, AAV Shuffle 100-3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM 10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.5O, AAVrh.43, AAVrh.62, AAVrh.48, AAVhu.19, AAVhu.ll, AAVhu.53, AAV4-8 / rh.64, AAVLG-9 / hu.39, AAV54.5 / hu.23, AAV54.2 / hu.22, AAV54.7 / hu.24, AAV54.1 / hu.21, AAV54.4R / hu.27,AAV46.2 / hu.28, AAV46.6 / hu.29, AAV128. l / hu.43, true type AAV (ttAAV), EGRENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7.10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-El, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV CHt-6. 1, AAV CHt- 6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-Pl, AAV CHt-P2,AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-Bl, AAV CKd-B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV CKd-B8, AAV CKd-Hl, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAV CKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-Fl, AAV CLg-F2, AAV CLg- F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1, AAV CLvl-1, AAV Clvl-10, AAV CLvl-2, AAV CLv-12, AAV CLvl-3, AAV CLv-1 3, AAV CLvl-4, AAV Clvl-7, AAV Clvl-8, AAV Clvl-9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-Dl, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-El, AAV CLv-Kl, AAV CLv-K3, AAV CLv- K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-Ml, AAV CLv-Ml 1, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-Rl, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV CSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8. 10, AAV CSp-8.2, AAV CSp- 8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1, AAVF11 / HSC11, AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5, AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, AAVF9 / HSC9, variants thereof, a hybrid or chimera of any of the foregoing AAV serotypes, or any combination thereof.80. The composition of any one of embodiments 75-79, wherein the AAV vector comprises an AAV variant engineered for reduced binding to heparin sulfate proteoglycans.81. The composition of any one of embodiments 75-80, wherein the AAV vector comprises a capsid protein comprising an insertion of a targeting peptide after amino acid 587 (AA587) of the AAV capsid protein or a functional equivalent of AA587, wherein the targeting peptide insertion results in enhanced transduction of target cells as compared to an AAV comprising a capsid protein that does not comprise the insertion.82. The composition of embodiment 81, wherein the targeting peptide is about seven amino acids in length.83. The composition of any one of embodiments 75-82, comprising a recombinant AAV (rAAV).84. The composition of any one of embodiments 1-83, further comprising one or more pharmaceutically acceptable excipients.85. The composition of any one of embodiments 1-84 for use in treating a disease or disorder.86. The composition of embodiment 85, wherein the gene is a therapeutic gene, and the disease or disorder is a retinal disease or disorder.87. A method of treating a disease or disorder in a subject, the method comprising: administering the composition of any one of claims 1-86 to the subject; and applying thermal energy to a target site of the subject sufficient to increase local temperature of the target site to an activating temperature, thereby inducing the expression of the gene product.88. The method of embodiment 87, wherein the composition is administered at a dose of about 1 x 109viral genomes to about 1.0 * 1012viral genomes.89. The method of embodiment 87, wherein the composition is administered at a dose of more than about 1.0 * 1012viral genomes.90. The method of any one of embodiments 87-89, wherein the composition is administered by intravitreal route, subretinal route, suprachoroidal route, intravenous route, intratumoral route, intracranial route, intramuscular route, intraarterial route, subcutaneous route, inhalation route, insufflation route, oral route, intranasal route, intraauricular route, or topical route.91. The method of any one of embodiments 87-90, wherein the method comprises a single administration of the composition to the subject.92. The method of any one of embodiments 87-91, wherein the method comprises administering the composition two or more times to the subject.93. The method of embodiment 92, wherein each of the two or more administrations are 1 month, 2 months, 6 months, 1 year, or more, apart.94. The method of any one of embodiments 87-93, wherein the period of time between the administering the composition and applying thermal energy is about 1 week, about 6 days, about 5 days, about 4 days, about 3 days, about 48 hours, about 44 hours, about 40 hours, about 35 hours, about 30 hours, about 25 hours, 20 hours, 15 hours, 10 hours, about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, about 10 minutes, or about 5 minutes, optionally the method comprises two or more applications of thermal energy, further optionally the period of time between administering the composition and applying thermal energy is the period of time between the administering the composition and the first application of thermal energy.95. The method of any one of embodiments 87-94, wherein the activating temperature is greater than 37°C.96. The method of embodiment 95, wherein the activating temperature comprises a temperature of at least 37.5° C to at most 70°C.97. The method of embodiment 96, wherein the activating temperature is about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, about 46.0°C, about 46.5°C, about 47.0°C, about 47.5°C, about 48.0°C, about 48.5 °C, about 49.0°C, about 49.5°C, about 50.0°C, about 50.5°C, about 51.0°C, about 51.5°C, about 52.0°C, about 52.5°C, about 53.0°C, about 53.5°C, about 54.0°C, about 54.5°C, about 55.0°C, about 55.5°C, about 56.0°C, about 56.5°C, about 57.0°C, about 57.5°C, about 58.0°C, about 58.5°C, about 59.0°C, about 59.5°C, about 60.0°C, about 60.5°C, about 61.0°C, about 61.5°C, about 62.0°C, about 62.5°C, about 63.0°C, about 63.5°C, about 64.0°C, about 64.5°C, about 65.0°C, about 65.5°C, about 66.0°C, about 66.5°C, about 67.0°C, about 67.3°C, about 67.5°C, about 68.0°C, about 68.5°C, about 69.0°C, about 69.5°C, or about 70.0°C; optionally, wherein the thermal energy is applied to the target site for a duration of time comprising about 1 second, about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, or about 1 minute.98. The method of any one of embodiments 96-97, wherein the activating temperature is about 37.5°C, about 38.0°C, about 38.5°C, about 39.0°C, about 39.5°C, about 40.0°C, about 40.5°C, about 41.0°C, about 41.5°C, about 42.0°C, about 42.5°C, about 43.0°C, about 43.5°C, about 44.0°C, about 44.5°C, about 45.0°C, about 45.5°C, or about 46.0°C; optionally, wherein the thermal energy is applied to the target site for a duration of time comprising about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes.99. The method of any one of embodiments 96-98, wherein the activating temperature comprises a temperature of about 42°C to about 43°C.100. The method of embodiment 99, wherein the activating temperature is about 42.0°C, about 42.1°C, about 42.2°C, about 42.3°C, about 42.4°C, about 42.5°C, about 42.6°C, about 42.7°C, about 42.8°C, about 42.9°C, or about 43.0°C.101. The method of any one of embodiments 87-100, wherein the thermal energy is applied to the target site for a duration of time comprising about 1 second to about 20 minutes.102. The method of embodiment 101, wherein the thermal energy is applied to the target site for a duration of time comprising about 1 second, about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes.103. The method of any one of embodiments 87-102, wherein the temperature at the target site is elevated to about 42°C to about 43°C after the applying; optionally, within at most 30 minutes from the applying.104. The method of embodiment 103, wherein the temperature at the target site is elevated to about 42°C to about 43 °C within 10 minutes or less after the applying.105. The method of any one of embodiments 103-104, wherein the temperature at the target site is elevated to about 42°C to about 43°C within about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, or about 10 minutes after the applying.106. The method of any one of embodiments 103-105, wherein the temperature at the target site is maintained at about 42°C to about 43°C for at least about 15 minutes to at least about 120 minutes.107. The method of any one of embodiments 103-106, wherein the temperature at the target site is maintained at 42°C to 43°C for at least 15 minutes, at least 20 minutes, at least 25 minutes, at least 30 minutes, at least 35 minutes, at least 40 minutes, at least 45 minutes, at least 50 minutes, at least 55 minutes, at least 60 minutes, at least 90 minutes, or at least 120 minutes.108. The method of embodiment 107, wherein the temperature at the target site is maintained at 42°C to 43 °C for at least 20 minutes.109. The method of embodiment 107, wherein the temperature at the target site is maintained at about 42°C to about 43 °C for about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, or about 20 minutes.110. The method of any one of embodiments 87-109, wherein applying thermal energy to the target site of the subject comprises the application of one or more of ciliary body hyperthermia, magnetic hyperthermia, microwave hyperthermia, inductive diathermy, contact hyperthermia, ultrasonic diathermy, focused ultrasound, and deep hyperthermia.111. The method of any one of embodiments 87-110, wherein applying thermal energy to the target site of the subject comprises use of a laser.112. The method of embodiment 111, wherein the laser is a 577 nm or a 532 nm laser.113. The method of any one of embodiments 111-112, wherein the laser is set at a power level of about 25% to about 40%.114. The method of any one of embodiments 111-112, wherein the laser is set at a power level of about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, or about 40%.115. The method of any one of embodiments 111-114, wherein the laser is set at a power level of about 30%.116. The method of any one of embodiments 111-115, wherein the laser is applied to the target site for a pulse of about 1 second to about 15 seconds.117. The method of any one of embodiments 111-116, wherein the laser is applied to the target site for a pulse of about 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 11 seconds, 12 seconds, 13 seconds, 14 seconds, or 15 seconds.118. The method of any one of embodiments 87-117, wherein upon applying thermal energy to the target site, the transcription of the gene, the transactivator gene, the recombinase gene, or any combination thereof, increases by at least 1.1 -fold.119. The method of any one of embodiments 87-118, wherein upon applying thermal energy to the target site, the levels of the gene product, the transactivator gene product, the recombinase gene product, or any combination thereof, increases by at least 1.1 -fold, wherein the gene product comprises RNA transcribed from the gene, optionally an mRNA, a protein translated from the mRNA, or both.120. The method of any one of embodiments 87-119, wherein the method comprises applying thermal energy to the subject two or more times.121. The method of embodiment 120, wherein each of the two or more applications of thermal energy are 1 week apart, 2 weeks apart, 3 weeks apart, 4 weeks apart, 5 weeks apart, or more.EXAMPLES

[0151] Some aspects of the embodiments discussed above are disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the present disclosure.Example 1 Expanded characterization of thermal switch activation

[0152] This Example provides data related to parameters for timing and temperatures for activating the thermal switches described herein.

[0153] Primary human T cells were transduced with a genetic circuit containing the S7YB thermal switch driving the expression of a luciferase reporter. Cells were heated at temperatures ranging from 37-67.3°C and heating durations from Is - 20 min. Expression of the Glue (glucose) reporter was measured by luminescence 24 hrs post-heating and is displayed in the 3D and 2D plots as shown in FIG. 3.

[0154] These data demonstrate that the thermal switch can be activated when heated at mild temperatures (~40-45°C) for durations in the order of minutes and at high temperatures (~45- 70°C) for short durations in the order of seconds.Example 2AAV-VEGFi treatment for Age-related Macular Degeneration

[0155] In wet age-regulated macular degeneration, atypical blood vessels appear around the macula and retina. These vessels can leak proteins or lipids, and they may also cause scarring. Anti -angiogenic therapies can be effective in treating diseases such as AMD. Vascular endothelial growth factor (VEGF) is the main protein that causes new blood vessels to form in the macula in wet AMD. VEGF inhibitors reduce leakage from blood vessels, prevent their growth, decrease swelling of the retina, reduce vision loss, and improve vision in wet AMD. A composition of the disclosure can be administered to a subject suffering from AMD via subretinal or intravitreal injection. pHSP-controlled or boosted VEGF inhibitor (VEGFi) biologic (e.g., antibody fragment, RTH258) can be delivered to the retina of the subject. The patients receive laser heating on a schedule or as needed to boost therapy.Example 3 AAV-C3i treatment of dry AMD

[0156] Dry AMD is a slow deterioration of the cells of the macula, often over many years, as the retinal cells die off and are not renewed. Without being bound by any particular theory, the complement system is thought to play a role in the progression of dry AMD. A composition ofthe disclosure for expressing an inhibitor of complement 3 (C3i) can be administered via subretinal or intravitreal injection. The methods disclosed herein will be used to drive, e.g., pHSP-controlled or boosted complement C3i expression in the retina. Patients can receive laser heating on a schedule or as needed to boost therapy.Example 4 AAV-immunomodulator for noninfectious uveitis

[0157] Uveitis is a chronic condition treated with steroid or systemic biologies (e.g., antiinflammatory). Constitutive unending treatment may lead to infection or tumorigenesis Any of the compositions disclosed herein comprising, e.g., AAV comprising a polynucleotide encoding a therapeutic gene products can be administered to a subject via subretinal or intravitreal injection. pHSP-controlled or pHSP-boosted cytokine / inhibitor (e.g., TNFi or IL-6i) can be induced upon thermal activation. Patients can receive heating (e.g., laser heating) on a schedule or as needed to boost therapy.Example 5AAV-delivered Thermally Induced Production of Anti-VEGF from ARPE-19 Cells

[0158] This Example provides data demonstrating AAV-delivered thermally induced production of anti-VEGF from retinal pigment epithelial cells employing the compositions and methods disclosed herein. FIGS. 7A-7D depict a non-limiting exemplary AAV circuit design (FIG. 7A), exemplary schematic (FIG. 7B), and data (FIGS. 7C-7D) related to studies thereof. AAV circuit design comprised of the S7YB synthetic thermal switch (TS) driving the thermally controlled expression of a VEGF inhibitor (RTH258) and EFl alpha promoter driving constitutive expression of a GFP reporter (FIG. 7A). Retinal epithelial cells (ARPE-19) were transduced using AAV2 with the TS circuit (FIG. 7B). Upon heating, ARPE-19 cells express RTH258 which inhibits VEGF activity. FIG. 7C depicts data related to the transduction of ARPE-19 cells at an MOI of 100,000 with AAV2 encoding for the TS.VEGFi, as measured by GFP expression by flow cytometry analysis. FIG. 7D depicts data related to the expression of VEGFi by AAV2-transduced (MOI 10,000) retinal epithelial cells at temperatures ranging from 37-45.1°C heated for a duration of 30 minutes.

[0159] In at least some of the previously described embodiments, one or more elements used in an embodiment can interchangeably be used in another embodiment unless such a replacement is not technically feasible. It will be appreciated by those skilled in the art that various other omissions, additions and modifications may be made to the methods and structures described above without departing from the scope of the claimed subject matter. All such modificationsand changes are intended to fall within the scope of the subject matter, as defined by the appended claims.

[0160] With respect to the use of substantially any plural and / or singular terms herein, those having skill in the art can translate from the plural to the singular and / or from the singular to the plural as is appropriate to the context and / or application. The various singular / plural permutations may be expressly set forth herein for sake of clarity. As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. Any reference to “or” herein is intended to encompass “and / or” unless otherwise stated.

[0161] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as “open” terms (e.g., the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases “at least one” and “one or more” to introduce claim recitations.However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles “a” or “an” limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases “one or more” or “at least one” and indefinite articles such as “a” or “an” (e.g., “a” and / or “an” should be interpreted to mean “at least one” or “one or more”); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the bare recitation of “two recitations,” without other modifiers, means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to “at least one of A, B, and C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “ a system having at least one of A, B, and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). In those instances where a convention analogous to “at least one of A, B, or C, etc.” is used, in general such a construction is intended in the sense one havingskill in the art would understand the convention (e.g., “ a system having at least one of A, B, or C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and / or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms.

[0162] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0163] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub-ranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 articles refers to groups having 1, 2, or 3 articles. Similarly, a group having 1-5 articles refers to groups having 1, 2, 3, 4, or 5 articles, and so forth.

[0164] While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those skilled in the art. The various aspects and embodiments disclosed herein are for purposes of illustration and are not intended to be limiting, with the true scope and spirit being indicated by the following claims.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A composition for treating a retinal disease or disorder in a subject, comprising: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product; wherein the first inducible promoter is capable of inducing transcription of the gene to generate the therapeutic gene product upon thermal stimulation.

2. A composition for treating a retinal disease or disorder in a subject comprising a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a transactivator gene, and a second promoter operably linked to a second polynucleotide comprising a therapeutic gene, wherein the first inducible promoter is capable of inducing transcription of the transactivator gene to generate a transactivator transcript in the presence of thermal stimulation, wherein the transactivator transcript is capable of being translated to generate a transactivator; and wherein, in the presence of the transactivator and; optionally, a transactivator-binding compound, the second promoter is capable of inducing transcription of a therapeutic gene to generate a therapeutic gene product.

3. A composition for treating a retinal disease or disorder in a subject, comprising: a viral vector comprising a nucleic acid comprising a first inducible promoter and a second promoter each operably linked to a first polynucleotide comprising a therapeutic gene and to a second polynucleotide comprising a transactivator gene, wherein the first inducible promoter is capable of inducing transcription of the therapeutic gene and the transactivator gene to generate a polycistronic transcript upon thermal stimulation, wherein, in the presence of the transactivator and a transactivator-binding compound, the second promoter is capable of inducing transcription of the therapeutic gene and the transactivator gene to generate a polycistronic transcript, and wherein the polycistronic transcript is capable of being translated to generate a transactivator and a therapeutic protein.

4. The composition of any one of claims 2-3, wherein the second promoter comprises one or more copies of a transactivator recognition sequence the transactivator is capable of binding to induce transcription, and wherein the transactivator is incapable of binding the transactivator recognition sequence in the absence of the transactivator-binding compound, optionally the one ormore copies of a transactivator recognition sequence comprise one or more copies of a tet operator (TetO).

5. The composition of any one of claims 2-4, wherein the transactivator comprises reverse tetracycline-controlled transactivator (rtTA).

6. The composition of any one of claims 2-4, wherein the transactivator comprises tetracycline-controlled transactivator (tTA).

7. The composition of any one of claims 2-6, wherein the transactivator-binding compound comprises tetracycline, doxycycline or a derivative thereof.

8. The composition of any one of claims 3-7, wherein the first polynucleotide and the second polynucleotide are operably linked to a tandem gene expression element, optionally the tandem gene expression element is an internal ribosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof.

9. A composition for treating a retinal disease or disorder in a subject, comprising: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a recombinase gene, wherein the first inducible promoter is capable of inducing transcription of the recombinase gene to generate a recombinase transcript upon thermal stimulation, and wherein the recombinase transcript is capable of being translated to generate a recombinase; a second promoter and a second polynucleotide comprising a therapeutic gene, wherein, in the absence of a recombination event, the second promoter and the second polynucleotide are not operably linked, wherein the recombinase is capable of catalyzing the recombination event, and wherein the second promoter and the second polynucleotide are operably linked after the recombination event such that the second promoter is capable of inducing transcription of the therapeutic gene to generate a therapeutic gene product.

10. The composition of claim 9, wherein the recombination event comprises removal of a sequence flanked by recombinase target sites or an inversion of a sequence flanked by recombinase target sites; optionally the second polynucleotide is flanked by recombinase target sites.

11. The composition of any one of claims 9-10, wherein, prior to the recombination event, the sequence of the therapeutic gene is inverted relative to the promoter.

12. The composition of any one of claims 9-11, comprising at least one stop cassette situated between the second promoter and the therapeutic gene, wherein the stop cassette comprises one or more stop sequences, and wherein the one or more stop cassettes are flanked by recombinase target sites.

13. The composition of any one of claims 1-12, wherein the therapeutic gene product is an RNA transcript and / or protein, optionally the transcript is an mRNA capable of being translated to generate the protein.

14. The composition of any one of claims 12-13, wherein the at least one stop cassette is configured to prevent transcription of the therapeutic gene and / or translation of the therapeutic transcript.

15. The composition of any one of claims 12-14, wherein the one or more stop sequences comprise a polyadenylation signal, a stop codon, a frame-shifting mutation, or any combination thereof.

16. The composition of any one of claims 2-15, wherein the second promoter comprises a ubiquitous or tissue-specific promoter.

17. The composition of any one of claims 2-16, wherein the second promoter comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focusforming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl l promoters from vaccinia virus, an elongation factor 1-alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3- phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P- kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CBH promoter, any variant thereof, or any combination thereof.

18. The composition of any one of claims 9-17, wherein the recombinase is Cre, Dre, Flp, KD, B2, B3, , HK022, HP1, y6, ParA, Tn3, Gin, <PC31, Bxbl, R4, derivatives thereof, or any combination thereof; optionally the recombinase is a Flp recombinase and the recombinase target sites are FRT sites or the recombinase is a Cre recombinase and the recombinase target sites are loxP sites.

19. The composition of any one of claims 1-18, wherein the first inducible promoter comprises a core promoter and at least one heat shock element (HSE).

20. The composition of claim 19, wherein the core promoter comprises the heat shock protein transcription start site of a gene selected from the group consisting of HSPA1A, HSPH1, HSPB1, HSPA6, and YB.

21. The composition of any one of claims 19-20, wherein the core promoter comprises the sequence of any one of SEQ ID NOs: 9-12 or a sequence having at least 85% identity to any one of SEQ ID NOs: 9-12.

22. The composition of any one of claims 19-20, wherein the core promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 9-12.

23. The composition of any one of claims 19-22, wherein the first inducible promoter comprises seven HSEs.

24. The composition of any one of claims 19-23, wherein each of the at least one HSE comprises the sequence of 5’ - nGAAnnTTCnnGAAn-3’.

25. The composition of any one of claims 19-24, wherein the at least one HSE comprises the sequence of any one of SEQ ID NOs: 1-8 or a sequence having at least 85% identity to any one of SEQ ID NOs: 1-8 or a sequence differing by one or two nucleotide mismatches relative to any one of SEQ ID NOs: 1-8.

26. The composition of any one of claims 19-24, wherein the at least one HSE comprises a sequence selected from the group consisting of SEQ ID NOs: 1-8.

27. The composition of any one of claims 1-18, wherein the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 13-20 or a sequence having at least 85% identity to any one of SEQ ID NOs: 13-20.

28. The composition of any one of claims 1-18, wherein the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 13-20.

29. The composition of any one of claims 1-18, wherein the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 21-34 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-34.

30. The composition of any one of claims 1-18, wherein the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-34.

31. A composition for treating a retinal disease or disorder in a subject, comprising: a viral vector comprising a nucleic acid comprising a leaky inducible promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product,-n-wherein the leaky inducible promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the leaky inducible promoter is capable of boosting transcription of the gene, thereby driving a second level of expression of the therapeutic gene product upon thermal stimulation.

32. The composition of claim 31, wherein the leaky inducible promoter comprises an HSPB, HSPB’2, HSP A / A, or HSP A / B promoter.

33. The composition of any one of claims 31-32, wherein the leaky inducible promoter comprises the sequence of any one of SEQ ID NOs: 21-24 and 28-29 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-24 and 28-29.

34. The composition of any one of claims 31-32, wherein the leaky inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-24 and 28-29.

35. A composition for treating a retinal disease or disorder in a subject, comprising: a viral vector comprising a nucleic acid comprising a tandem promoter operably linked to a polynucleotide comprising a gene encoding a therapeutic gene product, wherein the tandem promoter comprises a first inducible promoter and a second constitutive promoter, wherein the second constitutive promoter induces transcription of the gene in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the first inducible promoter is capable of boosting transcription of the gene upon thermal stimulation, thereby driving a second level of expression of the therapeutic gene product.

36. The composition of claim 35, wherein the tandem promoter comprises, from 5’ to 3’ : the first inducible promoter then the second constitutive promoter; or wherein the tandem promoter comprises, from 5’ to 3’ : the second constitutive promoter then the first inducible promoter.

37. A composition for treating a retinal disease or disorder in a subject, comprising: a viral vector comprising a nucleic acid comprising a first inducible promoter operably linked to a first polynucleotide comprising a gene encoding a therapeutic gene product and a second constitutive promoter operably linked to a second polynucleotide comprising the gene encoding a therapeutic gene productwherein the second constitutive promoter induces transcription of the gene encoding a therapeutic gene product from the second polynucleotide in the absence of thermal stimulation, thereby driving a first level of expression of the therapeutic gene product; and wherein the first inducible promoter is capable of inducing the transcription of the therapeutic gene from the first polynucleotide upon thermal stimulation, thereby boosting the expression of the therapeutic gene product to a second level of expression.

38. The composition of claim 37, wherein the nucleic acid comprises, from 5’ to 3’ : the first inducible promoter, the first polynucleotide, the second constitutive promoter, and the second polynucleotide; or wherein the nucleic acid comprises, from 5’ to 3’: the second constitutive promoter, the second polynucleotide, the first inducible promoter, and the first polynucleotide.

39. The composition of any one of claims 31-38, wherein the second level of expression is at least 1.1 -fold higher than the first level of expression.

40. The composition of any one of claims 31-39, wherein, after a first duration of time, the therapeutic gene product returns to the first level of expression from the second level of expression, wherein the first duration of time is about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks or more.

41. A composition for treating a retinal disease or disorder in a subject, comprising: a viral vector comprising a nucleic acid comprising a bidirectional promoter comprising a first inducible promoter in a first orientation, and a second constitutive promoter in a second orientation, wherein the first inducible promoter is operably linked to a first polynucleotide comprising a first gene encoding a therapeutic gene product, and wherein the second constitutive promoter is operably linked to a second polynucleotide comprising a second gene, wherein the first and second orientations are opposite of each other; wherein the second constitutive promoter is capable of inducing transcription of the second gene in the absence of thermal stimulation; and wherein the first inducible promoter is capable of inducing transcription of the first gene upon thermal stimulation to generate the therapeutic gene product.

42. The composition of claim 41, wherein the second gene encodes a therapeutic gene product; optionally, the therapeutic gene product is capable of inhibiting vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6); further optionally, the therapeutic gene product is selected from the group consisting of RPE65, Complement Factor 1 (CF1), CD59, FMS Related Receptor Tyrosine Kinase 1 (FLT-l), Rab escortprotein 1 (REP1), NADH hydrogenase 4 (ND4), MER proto-oncogene tyrosine kinase (MERTK), X-linked retinitis pigmentosa GTPase regulator (RPGR), or Retinoschisin (RSI).

43. The composition of any one of claims 35-41, wherein the second constitutive promoter comprises a ubiquitous or tissue-specific promoter.

44. The composition of any one of claims 35-42, wherein the second constitutive promoter comprises a promoter selected from the group consisting of a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), spleen focus-forming virus (SFFV), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl l promoters from vaccinia virus, an elongation factor 1-alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3 -phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), P-kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CBH promoter, or any combination thereof.

45. The composition of any one of claims 35-44, wherein the first inducible promoter comprises a core promoter and at least one heat shock element (HSE).

46. The composition of claim 45, wherein the core promoter comprises the heat shock protein transcription start site of a gene selected from the group consisting of HSPA1A, HSPH1, HSPB1, HSPA6, and YB.

47. The composition of any one of claims 45-46, wherein the core promoter comprises the sequence of any one of SEQ ID NOs: 9-12 or a sequence having at least 85% identity to any one of SEQ ID NOs: 9-12.

48. The composition of any one of claims 45-46, wherein the core promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 9-12.

49. The composition of any one of claims 45-48, wherein the first inducible promoter comprises seven HSEs.

50. The composition of any one of claims 45-49, wherein each of the at least one HSE comprises the sequence of 5’ - nGAAnnTTCnnGAAn-3’.

51. The composition of any one of claims 45-50, wherein the at least one HSE comprises the sequence of any one of SEQ ID NOs: 1-8 or a sequence having at least 85% identity to any one of SEQ ID NOs: 1-8 or a sequence differing by one or two nucleotide mismatches relative to any one of SEQ ID NOs: 1-8.

52. The composition of any one of claims 45-50, wherein the at least one HSE comprises a sequence selected from the group consisting of SEQ ID NOs: 1-8.

53. The composition of any one of claims 35-44, wherein the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 13-20 or a sequence having at least 85% identity to any one of SEQ ID NOs: 13-20.

54. The composition of any one of claims 35-44, wherein the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 13-20.

55. The composition of any one of claims 35-44, wherein the first inducible promoter comprises the sequence of any one of SEQ ID NOs: 21-34 or a sequence having at least 85% identity to any one of SEQ ID NOs: 21-34.

56. The composition of any one of claims 35-44, wherein the first inducible promoter comprises a sequence selected from the group consisting of SEQ ID NOs: 21-34.

57. The composition of any one of claims 1-56, wherein any two of the first inducible promoter, the second promoter, the leaky inducible promoter, the tandem promoter, the second constitutive promoter, and the bidirectional promoter are in opposite orientations or the same orientation relative to each other.

58. The composition of any one of claims 1-57, wherein the therapeutic gene product comprises an siRNA, an shRNA, a miRNA, or a protein.

59. The composition of any one of claims 1-58, wherein the therapeutic gene product is capable of inhibiting vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6).

60. The composition of claim 59, wherein the therapeutic gene product comprises an antibody or fragment thereof capable of binding to vascular endothelial growth factor (VEGF), Complement C3, tumor necrosis factor (TNF), or interleukin-6 (IL-6).

61. The composition of any one of claims 1-60, wherein the thermal stimulation comprises heating to an activating temperature, wherein the activating temperature is greater than 37°C.

62. The composition of claim 61, wherein the activating temperature comprises a temperature of at least 37°C to at most 70°C.

63. The composition of any one of claims 1-62, wherein the viral vector is or comprises an AAV vector, a lentivirus vector, a retrovirus vector, an adenovirus vector, a herpesvirus vector, a herpes simplex virus vector, a cytomegalovirus vector, a vaccinia virus vector, a MVA vector, a baculovirus vector, a vesicular stomatitis virus vector, a human papillomavirus vector, an avipox virus vector, a Sindbis virus vector, a VEE vector, a Measles virus vector, an influenza virus vector,a hepatitis B virus vector, an integration-deficient lentivirus (IDLV) vector derivatives thereof, or any combination thereof.

64. The composition of claim 63, wherein the viral vector is capable of integrating into a mammalian cell genome, and wherein the therapeutic gene product is capable of being expressed upon integration.

65. The composition of any one of claims 63-64, wherein the viral vector is a lentiviral vector, optionally human immunodeficiency virus 1 (HIV-1), human immunodeficiency virus 2 (HIV-2), visna-maedi virus (VMV) virus, caprine arthritis-encephalitis virus (CAEV), equine infectious anemia virus (EIAV), feline immunodeficiency virus (FIV), bovine immune deficiency virus (BIV), simian immunodeficiency virus (SIV), derivatives thereof, or any combination thereof.

66. The composition of any one of claims 1-65, wherein the viral vector comprises an AAV vector.

67. The composition of claim 66, wherein the AAV vector comprises a single-stranded AAV (ssAAV) vector or a self-complementary AAV (scAAV) vector.

68. The composition of any one of claims 66-67, wherein the AAV vector comprises AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, derivatives thereof, or any combination thereof.

69. The composition of any one of claims 67-68, wherein the AAV vector is or comprises an AAV selected from the group consisting of AAV9, AAV9 K449R (or K449R AAV9), AAV1, AAVrhlO, AAV-DJ, AAV-DJ8, AAV5, AAVPHP.B (PHP.B), AAVPHP.A (PHP. A), AAVG2B-26, AAVG2B-13, AAVTH1.1-32, AAVTH1.1-35, AAVPHP.B2 (PHP.B2), AAVPHP.B 3 (PHP.B3), AAVPHP.N / PHP.B-DGT, AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP, AAVPHP.B-ATT-T, AAVPHP.B-DGT-T, AAVPHP.B-GGT-T, AAVPHP.B- SGS, AAVPHP.B-AQP, AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B-SNP, AAVPHP.B-QGT, AAVPHP.B-NQT, AAVPHP.B- EGS, AAVPHP.B-SGN, AAVPHP.B-EGT, AAVPHP.B-DST, AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP, AAVPHP.B-SQP, AAVPHP.B-QLP, AAVPHP.B- TMP, AAVPHP.B-TTP, AAVPHP.S / G2A12, AAVG2 Al 5 / G2A3 (G2A3), AAVG2B4 (G2B4), AAVG2B5 (G2B5), PHP.S, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11, AAV 12, AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-lb, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa,AAV43-1, AAV43-12, AAV43-20, AAV43-21, AAV43-23, AAV43-25, AAV43-5, AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7, AAVl-7 / rh.48, AAVl-8 / rh.49, AAV2-15 / rh.62, AAV2-3 / rh.61, AAV2-4 / rh.5O, AAV2-5 / rh.51, AAV3. l / hu.6, AAV3. l / hu.9, AAV3-9 / rh.52, AAV3-1 l / rh.53, AAV4-8 / rl 1.64, AAV4-9 / rh.54, AAV4-19 / rh.55, AAV5-3 / rh.57, AAV5-22 / rh.58, AAV7.3 / hu.7, AAV16.8 / hu.lO, AAV16.12 / hu.ll, AAV29.3 / bb.l, AAV29.5 / bb.2, AAV106. l / hu.37, AAV1 14.3 / hu.4O, AAV127.2 / hu.41, AAV127.5 / hu.42, AAV128.3 / hu.44, AAV130.4 / hu.48, AAV145. l / hu.53, AAV145.5 / hu.54, AAV145.6 / hu.55, AAV161. 10 / hu.60, AAV161.6 / hu.61, AAV33. 12 / hu.l7, AAV33.4 / hu.l5, AAV33.8 / hu.l6, AAV52 / hu.l9, AAV52.1 / hu.2O, AAV58.2 / hu.25, AAVA3.3, AAVA3.4, AAVA3.5, AAVA3.7, AAVC1, AAVC2, AAVC5, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70, AAVpi.l, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu. 12, AAVH6, AAVH-l / hu.l, AAVH-5 / hu.3, AAVLG-10 / rh.40, AAVLG-4 / rh.38, AAVLG-9 / hu.39, AAVN721- 8 / rh.43, AAVCh.5, AAVCh.5Rl, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5Rl, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.l, AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.10, AAVhu.ll, AAVhu.13, AAVhu.15, AAVhu.16, AAVhu. 1 7, AAVhu.18, AAVhu.20, AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44Rl, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48Rl, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.l 4 / 9, AAVhu.t 19, AAVrh.2, AAVrh.2R, AAVrh.8, AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13, AAVrh.BR, AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49, AAVrh.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64Rl, AAVrh.64R2, AAVrh.67, AAVrh.73, AAVrh.74, AAVrh8R, AAVrh8R A586R mutant, AAVrh8R R533 A mutant, AAAV, BAAV, caprine AAV, bovine AAV, AAVhEl.l, AAVhErl.5, AAVhER1.14, AAVhErl.8, AAVhErl. 16, AAVhErl.18, AAVhErl.35, AAVhErl.7, AAVhErl.36, AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.3 1, AAVhEr2.36, AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01, AAV-LK02, AAV- LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV-LK09, AAV- LK10, AAV-LK11, AAV- LK12, AAV-LK13, AAV-LK14, AAV-LK15, AAV-LK16, AAV-LK17, AAV-LK18, AAV- LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV- PAEC11, AAV-PAEC12, AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV Shuffle 100-1, AAV Shuffle 100-3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM 10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.5O, AAVrh.43, AAVrh.62, AAVrh.48, AAVhu.19, AAVhu.ll, AAVhu.53, AAV4-8 / rh.64, AAVLG-9 / hu.39, AAV54.5 / hu.23, AAV54.2 / hu.22, AAV54.7 / hu.24, AAV54.1 / hu.21, AAV54.4R / hu.27,AAV46.2 / hu.28, AAV46.6 / hu.29, AAV128. l / hu.43, true type AAV (ttAAV), EGRENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7.10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-El, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV CHt-6. 1, AAV CHt- 6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-Pl, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-Bl, AAV CKd-B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV CKd-B8, AAV CKd-Hl, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAV CKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-Fl, AAV CLg-F2, AAV CLg- F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1, AAV CLvl-1, AAV Clvl-10, AAV CLvl-2, AAV CLv-12, AAV CLvl-3, AAV CLv-1 3, AAV CLvl-4, AAV Clvl-7, AAV Clvl-8, AAV Clvl-9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-Dl, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-El, AAV CLv-Kl, AAV CLv-K3, AAV CLv- K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-Ml, AAV CLv-Ml 1, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-Rl, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV CSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8. 10, AAV CSp-8.2, AAV CSp- 8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1, AAVF11 / HSC11, AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5,AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, AAVF9 / HSC9, variants thereof, a hybrid or chimera of any of the foregoing AAV serotypes, or any combination thereof.

70. The composition of any one of claims 1-69, further comprising one or more pharmaceutically acceptable excipients.

71. The composition of any one of claims 1-70 for use in treating a retinal disease or disorder.

72. A method of treating a retinal disease or disorder in a subject, the method comprising: administering the composition of any one of claims 1-71 to the subject; and applying thermal energy to a target site of the subject sufficient to increase local temperature of the target site to an activating temperature, thereby inducing the expression of the therapeutic gene product.

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