Fig. 5
From: Quantification of evolved DNA-editing enzymes at scale with DEQSeq
Validation of evolved Cas12f-ABE variants. a Agarose gels and b quantification of triple target base editing assay of the indicated clusters. The assays were performed with the same induction level as used in the screen. Highlighted with a box are variants that were further validated. Statistical results from t-tests comparing the variants to WT are included above the bars. c Sanger sequencing and d EditR quantification of E. coli gDNA base editing with indicated variants. Statistical results from t-tests comparing the variants to WT are included above the bars. e Schematic illustration of the GFP-to-BFP conversion. The employed sgRNA is underlined in black, with the protospacer-adjacent motif (PAM) sequence underlined in gray. The residues Y66 and L64 are marked in black. Bases that can be edited by the spABEs are marked in black. Two editing outcomes are depicted below with the residues and bases changed marked in black. f FACS profiles of HEK293T-EGFP reporter cells eight days after nucleofection with indicated ABE mRNAs in combination with the GFP-sgRNA-Y66H. c Quantification of GFP to BFP and GFP to no fluorescent protein (NoFP) edited cells. Statistical results from t-tests comparing the variants to WT are included above the bars. P-values: “ns” not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001