Some embodiments of the methods and compositions provided herein relate to systems comprising a promoter operably linked to a nucleic acid encoding a receptor, and an inducible promoter operably linked to a nucleic acid encoding a payload. In some embodiments, transcription from the inducible promoter is induced by activation of the receptor. In some embodiments, transcription from the inducible promoter is further modulated by inhibiting a signal between the activated receptor and the inducible promoter.

Aspects of the disclosure relate to compositions and methods for treating hereditary hearing loss and/or vision loss, for example, due to Usher syndrome, Type 3A. In some embodiments, the disclosure provides a recombinant adeno-associated virus comprising: (i) an AAV-S capsid protein, and (ii) an isolated nucleic acid comprising a transgene (e.g., a transgene for expressing a clarin-1 protein). The present disclosure also provides methods of treating hereditary hearing loss and/or vision loss (e.g., Usher Syndrome, Type 3A) using the same.

Methods are provided for altering target DNA in a cell genetically modified to express a Cas 9 enzyme that forms a co-localization complex with a guide RNA complementary to the target DNA and that cleaves the target DNA in a site specific manner. Methods include introducing into the cell a first foreign nucleic acid encoding a donor nucleic acid sequence, introducing into the cell from media surrounding the cell the guide RNA complementary to the target DNA and which guides the Cas 9 enzyme to the target DNA, wherein the RNA and the enzyme are members of a co-localization complex for the target DNA, wherein the donor nucleic acid sequence is expressed, wherein the guide RNA and the Cas 9 enzyme co-localize to the target DNA, the Cas 9 enzyme cleaves the target DNA and the donor nucleic acid is inserted into the target DNA to produce altered DNA in the cell.

The present invention relates to novel transposon constructs and uses thereof. The novel transposon constructs of this invention have been developed based on structure-guided engineering approaches of the IS608 transposon. Provided are polynucleotides encoding for transposon ends, which may advantageously be used for site-specific insertion of a nucleotide sequence of interest into the genome of a target cell or a target DNA molecule. The invention provides further nucleic acids, vectors and recombinant cells encoding or containing the improved polynucleotides encoding for transposon ends, as well as a transposase system. Hence, the invention provides many tools for molecular genetic approaches for genome alteration, such as cloning strategies. Furthermore provided are medical and non-medical uses of the polynucleotides of the invention. The invention is in particular useful as a tool for gene delivery in genetically modified cell based therapeutic approaches for treating various diseases and for genetic tagging of endogenous proteins in research.

This disclosure provides various TcBuster transposases and transposons, systems, and methods of use.

The present invention relates to a method for integrating one or more recombinant sites in a sole target gene of a cell genome comprising carrying out sequentially multiple site-specific recombination by using alternatively two different serine-integrases, wherein the selection marker is removed after each site-specific recombination by different transposases: An in vitro method for inserting at least one recombinant site in a target gene/locus from an isolated cell genome, said method comprising integrating a nucleotide sequence A into the target gene from an isolated cell genome by homologous recombination, wherein said nucleotide sequence A comprises a recombination cassette, flanked at 5′ and 3′ by target gene homology arms, and removing the selection marker by using the second transposase, thus obtaining a cell genome A comprising (i), (ii) and (iii) of nucleotide sequence A.

The present invention relates nucleic acid molecules and concatemers containing spacer sequences useful for the efficient packaging of viral particles so as to minimize the incorporation of contaminant nucleic acids into these vectors, as well as methods of producing such viral particles.

A system for controlling population of a biological species includes a population of genetically-modified individuals of the biological species, where both males and females in the genetically-modified population carry two mutations. The first mutation is a repressible genetic mutation that results in death of a juvenile individual of the first sex when the juvenile individual of the first sex comprises the repressible lethal mutation and is reared in the absence of a repressor or causes an individual to be sterile when reared in the absence of a repressor. The second mutation is an underdominant genetic mutation.

Methods and compositions for modifying the coding sequence of endogenous genes using rare-cutting endonucleases. The methods and compositions described herein can be used to modify the endogenous USH2A gene.

The invention relates to biotechnology, to gene therapy DNA vectors, a Escherichia coli strain JM110-NAS, a method of its production and selection of gene therapy DNA vector for targeted gene therapy. The method of a gene therapy DNA vector for targeted gene therapy includes construction of a 2408 bp vector containing a 688 bp replication origin, a 467 bp transcription terminator hGH-TA, a 137 bp regulatory region RNA-out of transposon TnlO, a 1018 bp kanamycin resistance gene, and a 68 bp poly linker. Vector cleaving by XhoI and BamHI restriction endonucleases and ligation with promoter and regulatory region, while a site containing the promoter region of a human is used for the production of the gene therapy DNA vector of interest. The kanamycin resistance gene is cleaved by SpeI restriction sites. The remaining fragment was ligated to itself. The invention allows high effective targeted gene therapy.