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Nat Methods. 2018 Nov;15(11):924-927. doi: 10.1038/s41592-018-0178-9. Epub 2018 Oct 30.

Engineered anti-CRISPR proteins for optogenetic control of CRISPR-Cas9.

Author information

1
Synthetic Biology Group, Institute for Pharmacy and Biotechnology (IPMB) and Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), University of Heidelberg, Heidelberg, Germany.
2
Department of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany.
3
Institute of Bioengineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
4
Swiss Institute of Bioinformatics (SIB), Lausanne, Switzerland.
5
Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany.
6
BioQuant Center and Cluster of Excellence CellNetworks, Heidelberg University, Heidelberg, Germany.
7
German Center for Infection Research (DZIF), Heidelberg, Germany.
8
Synthetic Biology Group, Institute for Pharmacy and Biotechnology (IPMB) and Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), University of Heidelberg, Heidelberg, Germany. roland.eils@bihealth.de.
9
Department of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany. roland.eils@bihealth.de.
10
Digital Health Center, Berlin Institute of Health (BIH) and Charité, Berlin, Germany. roland.eils@bihealth.de.
11
Health Data Science Unit, University Hospital Heidelberg, Heidelberg, Germany. roland.eils@bihealth.de.
12
Synthetic Biology Group, Institute for Pharmacy and Biotechnology (IPMB) and Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), University of Heidelberg, Heidelberg, Germany. dominik.niopek@bioquant.uni-heidelberg.de.
13
Department of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany. dominik.niopek@bioquant.uni-heidelberg.de.

Abstract

Anti-CRISPR proteins are powerful tools for CRISPR-Cas9 regulation; the ability to precisely modulate their activity could facilitate spatiotemporally confined genome perturbations and uncover fundamental aspects of CRISPR biology. We engineered optogenetic anti-CRISPR variants comprising hybrids of AcrIIA4, a potent Streptococcus pyogenes Cas9 inhibitor, and the LOV2 photosensor from Avena sativa. Coexpression of these proteins with CRISPR-Cas9 effectors enabled light-mediated genome and epigenome editing, and revealed rapid Cas9 genome targeting in human cells.

PMID:
30377362
DOI:
10.1038/s41592-018-0178-9

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