Figure 4.

Determination of the PAM Requirement of a Novel ScCas9-ABE Using the BEON Assay

(A) Samples of the reporter sequences consisting of the same S1-gRNA target but different PAM sequences (in blue). The target “A” base is highlighted in red in the bolded stop codon. (B) Representative fluorescence and bright-field (merged) images of HEK293 cells transfected with S1-gRNA, ScCas9-ABE, and S1NNG1 (AAG) or S1NNG2 (CAG) reporter plasmids, or the corresponding reporter alone. The PAM sequences are listed in yellow. Scale bars: 100 μm. (C) FACS quantification of GFP⁺ cells for various reporters with or without ScCas9-ABE editing. The PAM sequence is listed on each graph. ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.