Controllable genome editing with split-engineered base editors
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 a...
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Published in | Nature chemical biology Vol. 17; no. 12; pp. 1262 - 1270 |
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Main Authors | , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Nature Publishing Group
01.12.2021
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Subjects | |
Online Access | Get full text |
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Summary: | 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. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 AUTHOR CONTRIBUTIONS K.N.B., J.S., and R.M.K. conceived of the approach and designed the research. K.N.B., N.H.E., R.A.D., D.R., M.L., A.B., T.W., C.R.B., and A.M.G. performed experiments and analyzed data. K.N.B., N.H.E., and T.W., and Y.L. performed computational and statistical analysis. The manuscript was drafted by K.N.B., revised by N.H.E, J.S., and R.M.K., with added input and approval from all authors. |
ISSN: | 1552-4450 1552-4469 |
DOI: | 10.1038/s41589-021-00880-w |