Figure 1.

Screening of small molecules capable of improving prime editing

(A) Schematic diagram showing the screening strategy. The prime editor was designed to knock in a 40 bp DNA fragment into HEK3 site, and the resulting insertion can be amplified using PCR and quantified using gel electrophoresis. (B) Representative gel electrophoresis and ImageJ analysis of the effect of small molecules on the 40 bp knockin. Knockin bands were distinguishable from wild-type bands by gel electrophoresis (upper panel), and efficiency of the knockin was calculated using ImageJ software (lower panels). Nexturastat A, which improved knockin efficiency, is marked in red. (C) Summary of the effect of 54 small molecules on knockin. Each molecule was used at a concentration of 10 μM different drugs. The improvement ratios were calculated relative to DMSO. Three HDAC inhibitors, nexturastat A (DO1-f-2), abexinostat (DO1-d-4), and vorinostat (DO1-b-9), that significantly improved the efficiency of knockin are labeled by red boxes. Editing efficiencies were measured using gel electrophoresis and grayscale analysis.