A Split and Inducible Adenine Base Editor for Precise In Vivo Base Editing

Authors

Hongzhi Zeng
Qichen Yuan
Fei Peng
Dacheng Ma
Ananya Lingineni
Kelly Chee
Peretz Gilberd
Emmanuel C Osikpa
Zheng Sun
Xue Gao

Publication Date

9-11-2023

Journal

Nature Communications

DOI

10.1038/s41467-023-41331-5

PMID

37696818

PMCID

PMC10495389

PubMedCentral® Posted Date

9-11-2023

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Humans, Animals, Mice, Adenine, Proprotein Convertase 9, Gene Editing, Nucleotides, Molecular engineering, CRISPR-Cas9 genome editing, CRISPR-Cas systems

Abstract

DNA base editors use deaminases fused to a programmable DNA-binding protein for targeted nucleotide conversion. However, the most widely used TadA deaminases lack post-translational control in living cells. Here, we present a split adenine base editor (sABE) that utilizes chemically induced dimerization (CID) to control the catalytic activity of the deoxyadenosine deaminase TadA-8e. sABE shows high on-target editing activity comparable to the original ABE with TadA-8e (ABE8e) upon rapamycin induction while maintaining low background activity without induction. Importantly, sABE exhibits a narrower activity window on DNA and higher precision than ABE8e, with an improved single-to-double ratio of adenine editing and reduced genomic and transcriptomic off-target effects. sABE can achieve gene knockout through multiplex splice donor disruption in human cells. Furthermore, when delivered via dual adeno-associated virus vectors, sABE can efficiently convert a single A•T base pair to a G•C base pair on the PCSK9 gene in mouse liver, demonstrating in vivo CID-controlled DNA base editing. Thus, sABE enables precise control of base editing, which will have broad implications for basic research and in vivo therapeutic applications.

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