Fig. 6. Structure-guided engineering of Al3Cas12f furtherly improved editing efficiency.
a, Design of combinatorial mutants of Al3Cas12f. Engineered variants and codes are listed. b, Editing efficiency of 36 combinatorial mutants at SOD1 in K562 cells in comparison to Al3Cas12f wild type. The bar height represents the mean percentage of indels (n = 2 independent biological replicates). Individual data points are shown as dots. The dashed line indicates the level of editing efficiency of Al3Cas12f wild type. The lead K79R;M190K;E222K variant (no. 53, in red) is referred to as RKK. c, Comparison of editing efficiency between Al3Cas12f wild type (purple) and RKK (red) across eight TTTA-PAM-containing genomic loci in K562 cells. Data represent the mean indel percentage ± s.d. (n = 3 independent biological replicates). d, Box-and-whisker plot showing the data in c. The box outlines denote the first and third quartiles, the median is represented by the center line and the whiskers encompass the full data range. Individual data points are shown as dots. Significance was determined using a paired t-test.