Figure 2.

Characterization of HMD-G fusion to Cas9, multiple Cas9 variants, and other Cas9 orthologs in HEK293T cells

(A) Indel efficiencies of Cas9 and eeCas9 at 39 endogenous targets in HEK293T cells. (B) Summary of editing efficiencies of Cas9 and eeCas9 for targets at 41 endogenous targets (39 targets in A and two targets in D) in HEK293T cells. (C) Summary of target sites with an indel rate of over 50%. (D) Comparison of HDR-mediated precise editing efficiencies of Cas9 and eeCas9 in HEK293T cells. (E) Summary of editing efficiencies of Cas9 variants with or without HMG-D domain at multiple endogenous targets for non-NGG PAM in HEK293T cells. (F) Indel efficiencies of eCas9 1.1 and ee-eCas9 1.1 at 25 endogenous targets in HEK293T cells. (G) Summary of editing efficiencies of three high-fidelity Cas9 variants with or without HMG-D domain in HEK293T cells. (H) Indel efficiencies of CjCas9 and eeCjCas9 at eight endogenous targets in HEK293T cells. (I) Indel efficiencies of Nme2Cas9 and eeNme2Cas9 at six endogenous targets in HEK293T cells. Data in (A), (D), (F), (H), and (I) represent mean ± SD (n = 3 independent experiments). Because editing efficiencies were highly variable among all tested targets, statistical analysis of the average editing efficiencies including (E), (G), (H) (right). and (I) (right) were performed by using paired two-tailed Student's t test, while the remaining graphs were performed by using unpaired two-tailed Student's t test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.