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. 2020 Apr 16;28(7):1696–1705. doi: 10.1016/j.ymthe.2020.04.009

Figure 5.

Figure 5

Enrichment of Base-Edited Cells Using the BEON Assay

(A) Representative fluorescence and bright-field (merged) images of HEK293 cells transfected with the S1-reporter, S1-gRNA, and ABEmax before and after sorting. Scale bar: 100 μm. (B) BEAT analyses of the PCR amplicons of the endogenous site 1 for the cells before and 24 h (24h) after sorting. The red arrows point to the target “A” base at position 5. (C) Quantification of the A-to-G conversion efficiency of site 1 target for the sorted GFP-high and GFP-low HEK293 cells with the BEON reporter or a simple GFP-expressing construct. (D) Quantification of the A-to-G conversion efficiency of site 1 target for the sorted GFP-high and GFP-low HEK293 cells cultured for 1 more week. (E) Quantification of the A-to-G conversion efficiency of CAR target for the sorted GFP-high and GFP-low HEK293 cells using the BEON reporter. (F) Quantification of the A-to-G conversion efficiency of mouse Tmem5 target for the sorted GFP-high and GFP-low Neuro2A cells using the BEON reporter. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.