Generation Bio Co.
CLOSED-ENDED DNA (CEDNA) VECTORS FOR INSERTION OF TRANSGENES AT GENOMIC SAFE HARBORS (GSH) IN HUMANS AND MURINE GENOMES

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. Some ceDNA vectors further comprise cis-regulatory elements, including regulatory switches for regulation of the transgene expression after its insertion at a GSH