Fig. 4

Efficient adenine base editing mediated by SpRY-fused TadA8e monomer toward non-G PAMs in rice. a The gene construct of rBE62 used for adenine base editing in transgenic rice. Ubi-P, maize ubiquitin 1 promoter; TadA8e, the evolved E. coli TadA gene; XTEN, 16 amino acid flexible linker; NLS, nuclear localization signal. b–d Representative edited alleles of OsMPK13 (b) and OsGS1 (c), and OsGSK4 (d) generated by rBE62 in T0 transgenic rice lines. The PAM sequences for each target site are shown in the pale purple arrows; the candidate bases in the putative editing window for editing and detected nucleotide changes are highlighted in red and green, respectively; the nucleotide substitutions are underlined in the sequencing chromatograms. e–g Summary of nucleotide changes in the editing window of OsMPK13 (e), OsGS1 (f), and OsGSK4 (g) caused by rBE62 in T0 transgenic rice lines. The PAM sequences and the detected nucleotide changes are highlighted in green and red, respectively. h Summary of the genotyping results on T0 transgenic rice lines generated with rBE62. The PAM sequences and the target regions are highlighted in green and bold, respectively