Controllable genome editing with split-engineered base editors

Kiara N. Berríos, Niklaus H. Evitt, Rachel A. DeWeerd, Diqiu Ren, Meiqi Luo, Aleksia Barka, Tong Wang, Caroline R. Bartman, Yemin Lan, Abby M. Green, Junwei Shi, Rahul M. Kohli

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

DNA deaminase enzymes play key roles in immunity and have recently been harnessed for their biotechnological applications. In base editors (BEs), the combination of DNA deaminase mutator activity with CRISPR–Cas localization confers the powerful ability to directly convert one target DNA base into another. While efforts have been made to improve targeting efficiency and precision, all BEs so far use a constitutively active DNA deaminase. The absence of regulatory control over promiscuous deaminase activity remains a major limitation to accessing the widespread potential of BEs. Here, we reveal sites that permit splitting of DNA cytosine deaminases into two inactive fragments, whose reapproximation reconstitutes activity. These findings allow for the development of split-engineered BEs (seBEs), which newly enable small-molecule control over targeted mutator activity. We show that the seBE strategy facilitates robust regulated editing with BE scaffolds containing diverse deaminases, offering a generalizable solution for temporally controlling precision genome editing. [Figure not available: see fulltext.].

Original languageEnglish
Pages (from-to)1262-1270
Number of pages9
JournalNature Chemical Biology
Volume17
Issue number12
DOIs
StatePublished - Dec 2021

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