Multi-reporter selection for the design of active and more specific zinc-finger nucleases for genome editing.

ABSTRACT: Engineered nucleases have transformed biological research and offer great therapeutic potential by enabling the straightforward modification of desired genomic sequences. While many nuclease platforms have proven functional, all can produce unanticipated off-target lesions and have difficulty discriminating between homologous sequences, limiting their therapeutic application. Here we describe a multi-reporter selection system that allows the screening of large protein libraries to uncover variants able to discriminate between sequences with substantial homology. We have used this system to identify zinc-finger nucleases that exhibit high cleavage activity (up to 60% indels) at their targets within the CCR5 and HBB genes and strong discrimination against homologous sequences within CCR2 and

SUBMITTER: Oakes BL 

PROVIDER: S-EPMC4729830 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature