Figure 4.

Evaluation of SpCas9-Gem in Pluripotent Stem Cells

(A) Schematic summary of EGFP-to-BFP conversion targeting strategy. The location of the two amino acid changes and the Cas9 target site (red) are indicated.

(B and C) Representative flow cytometry plots (B) and graphical plot (C) depicting the proportion of BFP⁺ (targeting efficiency) and EGFP^neg cells (NHEJ frequency) assessed 3 days after the introduction of SpCas9 and SpCas9-Gem with and without a BFP homologous template in iPSCs constitutively expressing EGFP. Data represent the average ± SD of four independent experiments in two EGFP⁺⁺ iPSC lines. ^∗p < 0.0001, ^∗∗p < 0.000001.