TY - JOUR
T1 - A chemically controlled Cas9 switch enables temporal modulation of diverse effectors
AU - Wei, Cindy T.
AU - Popp, Nicholas A.
AU - Peleg, Omri
AU - Powell, Rachel L.
AU - Borenstein, Elhanan
AU - Maly, Dustin J.
AU - Fowler, Douglas M.
N1 - Publisher Copyright:
© 2023, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2023/8
Y1 - 2023/8
N2 - CRISPR–Cas9 has yielded a plethora of effectors, including targeted transcriptional activators, base editors and prime editors. Current approaches for inducibly modulating Cas9 activity lack temporal precision and require extensive screening and optimization. We describe a versatile, chemically controlled and rapidly activated single-component DNA-binding Cas9 switch, ciCas9, which we use to confer temporal control over seven Cas9 effectors, including two cytidine base editors, two adenine base editors, a dual base editor, a prime editor and a transcriptional activator. Using these temporally controlled effectors, we analyze base editing kinetics, showing that editing occurs within hours and that rapid early editing of nucleotides predicts eventual editing magnitude. We also reveal that editing at preferred nucleotides within target sites increases the frequency of bystander edits. Thus, the ciCas9 switch offers a simple, versatile approach to generating chemically controlled Cas9 effectors, informing future effector engineering and enabling precise temporal effector control for kinetic studies. [Figure not available: see fulltext.]
AB - CRISPR–Cas9 has yielded a plethora of effectors, including targeted transcriptional activators, base editors and prime editors. Current approaches for inducibly modulating Cas9 activity lack temporal precision and require extensive screening and optimization. We describe a versatile, chemically controlled and rapidly activated single-component DNA-binding Cas9 switch, ciCas9, which we use to confer temporal control over seven Cas9 effectors, including two cytidine base editors, two adenine base editors, a dual base editor, a prime editor and a transcriptional activator. Using these temporally controlled effectors, we analyze base editing kinetics, showing that editing occurs within hours and that rapid early editing of nucleotides predicts eventual editing magnitude. We also reveal that editing at preferred nucleotides within target sites increases the frequency of bystander edits. Thus, the ciCas9 switch offers a simple, versatile approach to generating chemically controlled Cas9 effectors, informing future effector engineering and enabling precise temporal effector control for kinetic studies. [Figure not available: see fulltext.]
UR - http://www.scopus.com/inward/record.url?scp=85149304636&partnerID=8YFLogxK
U2 - 10.1038/s41589-023-01278-6
DO - 10.1038/s41589-023-01278-6
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C2 - 36879061
AN - SCOPUS:85149304636
SN - 1552-4450
VL - 19
SP - 981
EP - 991
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 8
ER -