Activation of recombinases at specific DNA loci by zinc-finger domain insertions.

ABSTRACT: Recombinases have several potential advantages as genome editing tools compared to nucleases and other editing enzymes, but the process of engineering them to efficiently recombine predetermined DNA targets demands considerable investment of time and labor. Here we sought to harness zinc-finger DNA-binding domains (ZFDs) to program recombinase binding by developing fusions, in which ZFDs are inserted into recombinase coding sequences. By screening libraries of hybrid proteins, we optimized the insertion site, linker length, spacing and ZFD orientation and generated Cre-type recombinases that remain dormant unless the insertionally fused ZFD binds its target site placed in the vicinity of the recombinase binding site. The developed fusion improved targeted editing efficiencies of recombinas

SUBMITTER: Mukhametzyanova L 

PROVIDER: S-EPMC11631766 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature