Fig. 3
From: NmeCas9 is an intrinsically high-fidelity genome-editing platform
NmeCas9 and SpyCas9 have comparable editing efficiencies in human (HEK293T) cells when targeting the same chromosomal sites. a Western blot analysis of NmeCas9 and SpyCas9. HEK293T cells were transfected with the indicated Cas9 ortholog cloned in the same plasmid backbone and fused to the same HA epitope tags and NLSs. Top panel: anti-HA western blot (EP, empty sgRNA plasmid). Bottom panel: anti-GAPDH western blot, used as a loading control. Mobilities of protein markers are indicated. b T7E1 analysis of three previously validated SpyCas9 guides targeting the AAVS1 locus, in comparison with NmeCas9 guides targeting nearby AAVS1 sites (mean ± s.e.m., n = 3). c Representative T7EI analyses comparing editing efficiencies at the dual target sites DTS1, DTS3, DTS7, DTS8, and NTS7, using the indicated Cas9/sgRNA combinations. Products resulting from Cas9 genome editing are denoted by the red dots. d Quantitation of data from (c) (mean ± s.e.m., n = 3). Two-tailed paired Student’s T test showed significant difference between NmeCas9 and SpyCas9 editing of DTS1, DTS3, and DTS8 (p < 0.05)