Efficient generation of mouse models of human diseases via ABE- and BE-mediated base editing

TLDR

Adenine base editors (ABEs) can efficiently convert A to G, but their in vivo precision and efficiency remain to be fully characterized. The authors assessed ABE specificity by deep sequencing and whole‑genome sequencing of edited tissues. Microinjection of ABE mRNA and sgRNAs yielded up to 100% A‑to‑G conversion, producing mouse models with Ar and Hoxd13 mutations, and combined ABE with SaBE3 enabled simultaneous C‑to‑T and A‑to‑G edits, proving the approach’s high efficiency and precision for disease modeling and potential therapy.

Abstract

Abstract A recently developed adenine base editor (ABE) efficiently converts A to G and is potentially useful for clinical applications. However, its precision and efficiency in vivo remains to be addressed. Here we achieve A-to-G conversion in vivo at frequencies up to 100% by microinjection of ABE mRNA together with sgRNAs. We then generate mouse models harboring clinically relevant mutations at Ar and Hoxd13 , which recapitulates respective clinical defects. Furthermore, we achieve both C-to-T and A-to-G base editing by using a combination of ABE and SaBE3, thus creating mouse model harboring multiple mutations. We also demonstrate the specificity of ABE by deep sequencing and whole-genome sequencing (WGS). Taken together, ABE is highly efficient and precise in vivo, making it feasible to model and potentially cure relevant genetic diseases.