Fig. 3: Developing highly active and wide-window CBEs.

a Schematic diagram illustrating the stepwise movement of evoCDA insertion sites within FrCas9n. b Achievement of C-to-T editing activity at target sites by different CBEs (n = 4 biological replicates, data represent mean ± SD). c Comparing the editing activity of three CBEs (N-, ID822, and ID824) at two endogenous gene loci (n = 4 biological replicates, data represent mean ± SD). d The C-T conversion frequencies of each cytosine nucleotide were measured for ID824-evoCDA-FrCas9n at six endogenous gene loci, and the editing results were quantified and displayed in a heat map using EditR (n = 4 biological replicates, data represent mean ± SD). e Editing capability of N-evoCDA-FrCas9n and ID824-evoCDA-FrCas9n in 22-bp spacer region (n = 4 biological replicates, data represent mean ± SD). f Schematic illustrating the broad-range C-to-T editing induced by ID824-evoCDA-FrCas9n on both sides of a palindromic PAM. sgRNA1 targets C sites on the upstream sense strand, sgRNA2 targets C sites on the downstream antisense strand. g ID824-evoCDA-FrCas9n achieves ultra-wide-range editing across nearly 38 bases in the genome. C1–C22 represent target C sites on the sense strand upstream of the PAM, while G'1–G'22 represent the corresponding G sites on the complementary sense strand downstream of the PAM (n = 4 biological replicates, data represent mean ± SD).