Cas9 gRNA engineering for genome editing, activation and repression

Samira Kiani; Alejandro Chavez; Marcelle Tuttle; Richard N Hall; Raj Chari; Dmitry Ter-Ovanesyan; Jason Qian; Benjamin W Pruitt; Jacob Beal; Suhani Vora; Joanna Buchthal; Emma J K Kowal; Mohammad R Ebrahimkhani; James J Collins; Ron Weiss; George Church

doi:10.1038/NMETH.3580

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Cas9 gRNA engineering for genome editing, activation and repression

Journal article

Open access

Peer reviewed

Cas9 gRNA engineering for genome editing, activation and repression

Samira Kiani, Alejandro Chavez, Marcelle Tuttle, Richard N Hall, Raj Chari, Dmitry Ter-Ovanesyan, Jason Qian, Benjamin W Pruitt, Jacob Beal, Suhani Vora, …

Nature methods, Vol.12(11), pp.1051-1054

11/01/2015

DOI: 10.1038/NMETH.3580

PMCID: PMC4666719

PMID: 26344044

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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.

Flow Cytometry

Mutagenesis

Mutation

Binding Sites

CRISPR-Associated Proteins - genetics

CRISPR-Cas Systems - genetics

Fluorescent Dyes - analysis

Gene Deletion

Genes, Reporter

Genetic Engineering - methods

Genetic Vectors

Genome

HEK293 Cells

Humans

Microscopy, Fluorescence

RNA Editing

RNA, Guide, CRISPR-Cas Systems

Transcription, Genetic

Details

Title: Subtitle: Cas9 gRNA engineering for genome editing, activation and repression
Creators: Samira Kiani - Massachusetts Institute of Technology
Alejandro Chavez - Massachusetts General Hospital
Marcelle Tuttle - Harvard University
Richard N Hall - Massachusetts Institute of Technology
Raj Chari - Harvard University
Dmitry Ter-Ovanesyan - Harvard Medical School
Jason Qian - Harvard University
Benjamin W Pruitt - Harvard University
Jacob Beal - RTX (United States)
Suhani Vora - Harvard University
Joanna Buchthal - Harvard University
Emma J K Kowal - Harvard University
Mohammad R Ebrahimkhani - Emergent Behaviors of Integrated Cellular Systems
James J Collins - Massachusetts Institute of Technology
Ron Weiss - Massachusetts Institute of Technology
George Church - Harvard University
Resource Type: Journal article
Publication Details: Nature methods, Vol.12(11), pp.1051-1054
DOI: 10.1038/NMETH.3580
PMID: 26344044
PMCID: PMC4666719
NLM abbreviation: Nat Methods
ISSN: 1548-7091
eISSN: 1548-7105
Grant note: Canadian Institutes of Health Research P50 HG005550 / NHGRI NIH HHS R01 CA155320 / NCI NIH HHS 5R01CA155320-04 / NCI NIH HHS P50 GM098792 / NIGMS NIH HHS T32 CA009216 / NCI NIH HHS 5T32CA009216-34 / NCI NIH HHS T32 GM007598 / NIGMS NIH HHS
Language: English
Date published: 11/01/2015
Academic Unit: Electrical and Computer Engineering
Record Identifier: 9984627188902771

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