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Nat Methods. 2015 Nov;12(11):1051-4. doi: 10.1038/nmeth.3580. Epub 2015 Sep 7.

Cas9 gRNA engineering for genome editing, activation and repression.

Author information

1
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
2
Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
3
Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.
4
Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA.
5
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
6
Raytheon BBN Technologies, Cambridge, Massachusetts, USA.
7
Center for Emergent Behaviors of Integrated Cellular Systems (EBICS), Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
8
Institute for Medical Engineering &Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
9
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
10
Harvard-MIT Program in Health Sciences and Technology, Cambridge, Massachusetts, USA.

Abstract

We demonstrate that by altering the length of Cas9-associated guide RNA (gRNA) we were able to control Cas9 nuclease activity and simultaneously perform genome editing and transcriptional regulation with a single Cas9 protein. We exploited these principles to engineer mammalian synthetic circuits with combined transcriptional regulation and kill functions governed by a single multifunctional Cas9 protein.

PMID:
26344044
PMCID:
PMC4666719
DOI:
10.1038/nmeth.3580
[Indexed for MEDLINE]
Free PMC Article

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