Fig. 2: Mb2Cas12a and engineered Cas12a variants broaden the genome editing scope in rice.

a Targeted mutagenesis efficiencies (percentage) of ten Cas12a orthologs at two VTTV PAM sites in rice protoplasts. WT, protoplasts transformed with water. b Targeted mutagenesis efficiencies (percentage) of Mb2Cas12a at 18 VTTV PAM sites in rice protoplasts. c Summary of editing and biallelic editing efficiencies by Mb2Cas12a at two VTTV PAM sites in transgenic rice T₀ lines. d Comparison of editing efficiencies (percentage) of five Cas12a RVR variants at six TATV PAM sites in rice protoplasts. Treatments with the same letter are not significantly different when α = 0.05 by Tukey’s Honest Significant Difference test (two-sided). e A box plot showing targeted mutagenesis efficiencies (percentage) of Mb2Cas12a-RVRR variant at 51 canonical and altered PAM sites in rice protoplasts. V = A, C, and G. Y = T and C. S = C and G. R = A and G. Each data point represents the mean of three biological replicates (n = 3). The middle line in each box denotes the median. The upper and lower bounds of each box denote the first quartile (25%) and the third quartile (75%), respectively. The top and bottom of whiskers denote the maxima and minima, respectively. f Genome-wide analysis of targetable PAM sites by Mb2Cas12a-RVRR variant in rice (Oryza sativa). g and h, Insertion and deletion (Indel) frequencies (percentage) of four Cas12a orthologs at two TTTV PAM sites at 32 and 22 °C in rice protoplasts. One asterisk (p < 0.05) and two asterisks (p < 0.01) indicate significant differences between two treatments using two-sided Student’s t-test. p = 0.000583 (Er vs. Mb2); p = 0.000456 (Lb5 vs. Mb2); p = 0.012092 (Bs vs. Mb2). i Indel frequencies (percentage) of Mb2Cas12a at two VTTV PAM sites at 32 and 22 °C in rice protoplasts. Data in a, b, d, g–i are presented as mean values ± SEM. n = 3 biologically independent samples. Source data are provided as a Source Data file.