Supplementary Figure 4: Optimizing the coding sequence of high-fidelity and PAM variant Cas9 enzymes improves protein expression
From: Optimized base editors enable efficient editing in cells, organoids and mice
a. Giemsa stained NIH/3T3 cells following transduction with P2A-Puro lentiviruses, as indicated, and selection in puro for 6 days. Experiment was repeated 3 times with similar results. b. Flow cytometry plots showing fluorescence of GFP linked to original and optimized HF1, PAM variant, and BE3 enzymes. While most cells expressing optimized versions show much higher GFP fluorescence, a small fraction show low levels of GFP expression. This is likely due to integration-site specific effects on EF1-mediated transcription. c. Quantitation of mean GFP fluorescence intensity from original and optimized HF1, PAM variant, and BE3 enzymes. Error bars represent s.e.m., n = 3 biologically independent experiments.
