A Novel Cas9 Mutant Confers Reduced Off-Target Gene Editing While Maintaining on-Target Potency

We will present the screening of a Cas9 variant with increased specificity from a library of 250,000 recombinant clones. We will demonstrate robust transfection efficiency by lipofection and electroporation as RiboNucleoProtein (RNP) in various established or primary cell lines.

We will verify that this Cas9 mutant combines strongly reduced off-target editing with maintained on-target activity, compared with the Wild-type S.p Cas9 nuclease, when used as RNP. We will review high efficiency Knock-In applications with both short (<120 nt) single-stranded donor oligonucleotides and long (up to 2kb) single-stranded donor DNA fragments.