Fig. 4
The endonuclease activity assays of purified Cas RNPs. a In vitro cleavage on the target plasmid I (single cleavage site) by Cas9 RNPs or CL7–Cas9 RNPs, as well as on the target plasmid II (two cleavage sites) by Cas12a RNPs. b Delivery of Cas9 RNPs and donor ssDNA in BFP-HEK293 cells can induce HDR-mediated genome editing which can convert them into GFP-HEK 293 cells. c The bright-filed and fluorescent images of BFP-HEK293 cells after delivery of Cas9 RNPs (left), donor ssDNA only (middle), and Cas9 RNPs together with ssDNA donor (right) by lipofectamine CRISPRMAX in 48 h later. Scale bars: 100 μM. d The HDR efficiency was determined by GFP expression due to BFP editing according to the flow cytometry data (Supplementary Figure 8), including the prepared Cas9 RNPs in this work with ssDNA donor (red), ssDNA donor only (blue), and Cas9 enzyme). Data are shown as the mean ± SD
