Abstract: The present disclosure provides compositions and methods of restoring or enhancing visual function in an individual by administering to the individual a pharmaceutical composition comprising a recombinant adeno-associated viral (rAAV) vector having a polynucleotide sequence that encodes a medium wavelength cone opsin (MW-opsin). The MW-opsin is expressed in a retinal cell in the individual, thereby restoring or enhancing visual function.

Abstract: The application describes ceDNA vectors having linear and continuous structure for gene editing. ceDNA vectors comprise an expression cassette flanked by two ITR sequences, where the expression cassette encodes a gene editing molecule. Some ceDNA vectors further comprise cis-regulatory elements, including regulatory switches. Further provided herein are methods and cell lines for reliable gene editing using the ceDNA vectors.

Abstract: Aspects of the disclosure relate to a nucleic acid comprising a heterologous nucleic acid insert flanked by interrupted self-complementary sequences, wherein one self-complementary sequence is interrupted by a cross-arm sequence forming two opposing, lengthwise-symmetric stem-loops, and wherein the other of the self-complementary sequences is interrupted by a truncated cross-arm sequence. Methods of delivering the nucleic acid to a cell are also provided.

Abstract: Provided herein are methods and constructs comprising close-ended DNA (ceDNA vectors) for maintaining or sustaining a level of transgene expression at a predetermined level or range for a predefined time, or increasing the level of transgene expression in a cell or a subject, where the transgene expression level can be modulated (e.g., increased) with one or more subsequent administrations (e.g., a re-dose or a booster administration) after an initial priming administration. Provided are methods for personalizing gene therapy throughout an individuals' lifespan to express a transgene at a level that meets an individual's needs, by modulating expression levels of a transgene expressed by ceDNA vector incrementally, or in a step-by-step manner, with one or more administrations after an initial priming administration (e.g., at time 0), thereby enabling titration of the level of expression of the transgene to a desired predetermined expression level or to a desired expression level range.

Abstract: Provided herein are methods and constructs related to minimizing immune responses using inhibitors of the immune response, in particular the innate immune response, when administering a desired transgene in a cell achieved by delivery of the transgene with repeated doses of a ceDNA vector.

Abstract: Aspects of the disclosure relate to a nucleic acid comprising a heterologous nucleic acid insert flanked by interrupted self-complementary sequences, wherein one self-complementary sequence is interrupted by a cross-arm sequence forming two opposing, lengthwise-symmetric stem-loops, and wherein the other of the self-complementary sequences is interrupted by a truncated cross-arm sequence. Methods of delivering the nucleic acid to a cell are also provided.

Abstract: Aspects of the disclosure relate to a nucleic acid comprising a heterologous nucleic acid insert flanked by interrupted self-complementary sequences, wherein one self-complementary sequence is interrupted by a cross-arm sequence forming two opposing, lengthwise-symmetric stem-loops, and wherein the other of the self-complementary sequences is interrupted by a truncated cross-arm sequence. Methods of delivering the nucleic acid to a cell are also provided.

Abstract: The application describes ceDNA vectors having linear and continuous structure for insertion of a transgene into a gene safe harbor (GSH) in a genome, e.g., mammalian genome. ceDNA vectors can comprise at least one ITR sequence, or two ITR sequences, a transgene, and at least one nucleic acid sequence that specifically binds to, or hybridizes to a GSH locus. Some ceDNA vectors comprise at least one GSH homology arm (GSH HA), e.g., a 5? GSH HA, and/or a 3? GSH HA, and some ceDNA vectors comprise a guide RNA (gRNA) or guide DNA (gDNA) that specifically targets a region in the GSH locus and/or a 5? or 3? GSH HA herein. Some ceDNA vectors also comprise a gene editing cassette that encodes a gene editing molecule.

Abstract: The technology described herein relates to methods, compositions and in silico screening approaches for identifying and validating genomic safe harbors (GSHs) in mammalian genomes, including human genomes. Another aspects relates to recombinant nucleic acid vectors, including non-viral and viral vectors comprising a portion of the GSH loci, or gRNA sequences specific to a GSH loci, and methods for use of the vectors for insertion of a gene of interest into a GSH loci.

Abstract: Aspects of the disclosure relate to a nucleic acid comprising a heterologous nucleic acid insert flanked by interrupted self-complementary sequences, wherein one self-complementary sequence is interrupted by a cross-arm sequence forming two opposing, lengthwise-symmetric stem-loops, and wherein the other of the self-complementary sequences is interrupted by a truncated cross-arm sequence. Methods of delivering the nucleic acid to a cell are also provided.

Abstract: The present invention provides an adeno-associated virus 4 (AAV4) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV4 vectors and particles.

Abstract: The present invention provides an adeno-associated virus 4 (AAV4) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV4 vectors and particles.

Abstract: The present invention provides an adeno-associated virus 4 (AAV4) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV4 vectors and particles.

Abstract: The present invention provides an adeno-associated virus 4 (AAV4) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV4 vectors and particles.

Abstract: The present invention provides an adeno-associated virus 4 (AAV4) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV4 vectors and particles.

Abstract: The present invention provides an adeno-associated virus 4 (AAV4) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV4 vectors and particles.

Abstract: The present invention provides an adeno-associated virus 5 (AAV5) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV5 vectors and particles.

Abstract: A method of producing an adeno-associated virus (AAV) in an insect cell comprising (i) providing at least one insect cell-compatible vector comprising a first nucleotide sequence comprising at least one AAV ITR nucleotide sequence, a second nucleotide sequence containing an open reading frame encoding AAV VP1, VP2, and VP3 capsid proteins, a third nucleotide sequence comprising a Rep52 or a Rep40 coding sequence, and a fourth nucleotide sequence comprising a Rep78 or a Rep68 coding sequence, (ii) introducing the at least one insect cell-compatible vector into an insect cell, and (iii) maintaining the insect cell under conditions such that AAV is produced. Also provided are recombinant AAV made in accordance with the method, insect cell-compatible vectors, and insect cells comprising nucleotide sequences for production of AAV in an insect cell.

Abstract: The present invention provides an adeno-associated virus 5 (AAV5) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV5 vectors and particles.

Abstract: The present invention provides methods of delivering nucleic acids to specific regions, tissues and cell types of the CNS. More particularly the invention provides methods of delivering nucleic acids to cells of the CNS such as cerebellar cells and ependymal cells. The invention also provides methods of delivering nucleic acids to cells of the lung such as alveolar cells using AAV5 vectors and particles.