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Title: Cas9 gRNA engineering for genome editing, activation and repression

Here 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.
Authors:
 [1] ;  [2] ;  [3] ;  [1] ;  [4] ;  [5] ;  [5] ;  [3] ; ORCiD logo [6] ; ORCiD logo [7] ;  [3] ;  [3] ;  [1] ;  [8] ;  [1] ;  [5]
  1. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
  2. Harvard Univ., Cambridge, MA (United States); Massachusetts Hospital, Boston, MA (United States); Harvard Medical School, Boston, MA (United States)
  3. Harvard Univ., Cambridge, MA (United States)
  4. Harvard Medical School, Boston, MA (United States)
  5. Harvard Univ., Cambridge, MA (United States); Harvard Medical School, Boston, MA (United States)
  6. Raytheon BBN Technologies, Cambridge, MA (United States)
  7. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Harvard Univ., Cambridge, MA (United States); Harvard Medical School, Boston, MA (United States)
  8. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Harvard Univ., Cambridge, MA (United States); Broad Institute of MIT and Harvard, Cambridge, MA (United States); Harvard-MIT Program in Health Sciences and Technology, Cambridge, MA (United States)
Publication Date:
Grant/Contract Number:
FG02-02ER63445
Type:
Accepted Manuscript
Journal Name:
Nature Methods
Additional Journal Information:
Journal Volume: 12; Journal Issue: 11; Journal ID: ISSN 1548-7091
Publisher:
Nature Publishing Group
Research Org:
Harvard Univ., Cambridge, MA (United States)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1347131

Kiani, Samira, Chavez, Alejandro, Tuttle, Marcelle, Hall, Richard N., Chari, Raj, Ter-Ovanesyan, Dmitry, Qian, Jason, Pruitt, Benjamin W., Beal, Jacob, Vora, Suhani, Buchthal, Joanna, Kowal, Emma J. K., Ebrahimkhani, Mohammad R., Collins, James J., Weiss, Ron, and Church, George. Cas9 gRNA engineering for genome editing, activation and repression. United States: N. p., Web. doi:10.1038/nmeth.3580.
Kiani, Samira, Chavez, Alejandro, Tuttle, Marcelle, Hall, Richard N., Chari, Raj, Ter-Ovanesyan, Dmitry, Qian, Jason, Pruitt, Benjamin W., Beal, Jacob, Vora, Suhani, Buchthal, Joanna, Kowal, Emma J. K., Ebrahimkhani, Mohammad R., Collins, James J., Weiss, Ron, & Church, George. Cas9 gRNA engineering for genome editing, activation and repression. United States. doi:10.1038/nmeth.3580.
Kiani, Samira, Chavez, Alejandro, Tuttle, Marcelle, Hall, Richard N., Chari, Raj, Ter-Ovanesyan, Dmitry, Qian, Jason, Pruitt, Benjamin W., Beal, Jacob, Vora, Suhani, Buchthal, Joanna, Kowal, Emma J. K., Ebrahimkhani, Mohammad R., Collins, James J., Weiss, Ron, and Church, George. 2015. "Cas9 gRNA engineering for genome editing, activation and repression". United States. doi:10.1038/nmeth.3580. https://www.osti.gov/servlets/purl/1347131.
@article{osti_1347131,
title = {Cas9 gRNA engineering for genome editing, activation and repression},
author = {Kiani, Samira and Chavez, Alejandro and Tuttle, Marcelle and Hall, Richard N. and Chari, Raj and Ter-Ovanesyan, Dmitry and Qian, Jason and Pruitt, Benjamin W. and Beal, Jacob and Vora, Suhani and Buchthal, Joanna and Kowal, Emma J. K. and Ebrahimkhani, Mohammad R. and Collins, James J. and Weiss, Ron and Church, George},
abstractNote = {Here 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.},
doi = {10.1038/nmeth.3580},
journal = {Nature Methods},
number = 11,
volume = 12,
place = {United States},
year = {2015},
month = {9}
}