A novel base editor SpRY-ABE8eF148A mediates efficient A-to-G base editing with a reduced off-target effect

Adenine base editors (ABEs) can mediate two transition mutations, A-to-G and T-to-C, which are suitable for repairing G & BULL;C-to-T & BULL;A pathogenic variants, the most significant human pathogenic variant. By combining the protospacer adjacent motif (PAM)less SpRY nuclease with F148A-mutated TadA*8e deaminase, we developed a new editor, SpRY-ABE8eF148A, in this study, which has narrowed the editing range and enhanced A-to-G editing efficiency in most sites with NR/YN PAMs. Furthermore, compared with SpRY-ABE8e, SpRY-ABE8eF148A significantly decreased the RNA off-target effect. Therefore, this engineered base editor, SpRY-ABE8eF148A, expanded the editing scope and improved the editing precision for G & BULL;C-to-T & BULL;A pathogenic variants. Besides, we established a bioinformatics tool, adenine base-repairing sgRNA database of pathogenic variant (ARDPM), to facilitate the development of precise editors.

Automated summary

We developed a new editor, SpRY-ABE8eF148A, which can mediate A-to-G and T-to-C transition mutations, suitable for repairing the most significant G & BULL;C-to-T & BULL;A pathogenic variants. Compared with SpRY-ABE8e, SpRY-ABE8eF148A narrows the editing range and enhances A-to-G editing efficiency in most sites with NR/YN PAMs. Furthermore, it significantly decreases the RNA off-target effect.