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Title: Highly efficient Cas9-mediated transcriptional programming

Abstract

The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. Here we describe an improved transcriptional regulator through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. Here, we demonstrate its utility in activating endogenous coding and non-coding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).

Authors:
 [1];  [2];  [3];  [4];  [4];  [2];  [5];  [6];  [4];  [4];  [2];  [4];  [2];  [7];  [7];  [6];  [7];  [8];  [2]
  1. Harvard Univ., Cambridge, MA (United States); Massachusetts General Hospital, Boston, MA (United States); Harvard Medical School, Boston, MA (United States)
  2. Harvard Univ., Cambridge, MA (United States); Harvard Medical School, Boston, MA (United States)
  3. Harvard Univ., Cambridge, MA (United States); Harvard Medical School, Boston, MA (United States); Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
  4. Harvard Univ., Cambridge, MA (United States)
  5. Harvard Medical School, Boston, MA (United States); Tsinghua Univ., Beijing (China)
  6. Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
  7. Harvard Medical School, Boston, MA (United States)
  8. Harvard Univ., Cambridge, MA (United States); Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
Publication Date:
Research Org.:
Harvard Univ., Cambridge, MA (United States)
Sponsoring Org.:
USDOE; U.S. National Institutes of Health (NIH); National Cancer Institute (NCI); National Science Foundation (NSF)
OSTI Identifier:
1347133
Grant/Contract Number:
FG02-02ER63445; 5T32CA009216-34; P50 HG005550
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Nature Methods
Additional Journal Information:
Journal Volume: 12; Journal Issue: 4; Journal ID: ISSN 1548-7091
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Chavez, Alejandro, Scheiman, Jonathan, Vora, Suhani, Pruitt, Benjamin W., Tuttle, Marcelle, Iyer, Eswar P. R., Lin, Shuailiang, Kiani, Samira, Guzman, Christopher D., Wiegand, Daniel J., Ter-Ovanesyan, Dmitry, Braff, Jonathan L., Davidsohn, Noah, Housden, Benjamin E., Perrimon, Norbert, Weiss, Ron, Aach, John, Collins, James J., and Church, George M. Highly efficient Cas9-mediated transcriptional programming. United States: N. p., 2015. Web. doi:10.1038/nmeth.3312.
Chavez, Alejandro, Scheiman, Jonathan, Vora, Suhani, Pruitt, Benjamin W., Tuttle, Marcelle, Iyer, Eswar P. R., Lin, Shuailiang, Kiani, Samira, Guzman, Christopher D., Wiegand, Daniel J., Ter-Ovanesyan, Dmitry, Braff, Jonathan L., Davidsohn, Noah, Housden, Benjamin E., Perrimon, Norbert, Weiss, Ron, Aach, John, Collins, James J., & Church, George M. Highly efficient Cas9-mediated transcriptional programming. United States. doi:10.1038/nmeth.3312.
Chavez, Alejandro, Scheiman, Jonathan, Vora, Suhani, Pruitt, Benjamin W., Tuttle, Marcelle, Iyer, Eswar P. R., Lin, Shuailiang, Kiani, Samira, Guzman, Christopher D., Wiegand, Daniel J., Ter-Ovanesyan, Dmitry, Braff, Jonathan L., Davidsohn, Noah, Housden, Benjamin E., Perrimon, Norbert, Weiss, Ron, Aach, John, Collins, James J., and Church, George M. Mon . "Highly efficient Cas9-mediated transcriptional programming". United States. doi:10.1038/nmeth.3312. https://www.osti.gov/servlets/purl/1347133.
@article{osti_1347133,
title = {Highly efficient Cas9-mediated transcriptional programming},
author = {Chavez, Alejandro and Scheiman, Jonathan and Vora, Suhani and Pruitt, Benjamin W. and Tuttle, Marcelle and Iyer, Eswar P. R. and Lin, Shuailiang and Kiani, Samira and Guzman, Christopher D. and Wiegand, Daniel J. and Ter-Ovanesyan, Dmitry and Braff, Jonathan L. and Davidsohn, Noah and Housden, Benjamin E. and Perrimon, Norbert and Weiss, Ron and Aach, John and Collins, James J. and Church, George M.},
abstractNote = {The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. Here we describe an improved transcriptional regulator through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. Here, we demonstrate its utility in activating endogenous coding and non-coding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).},
doi = {10.1038/nmeth.3312},
journal = {Nature Methods},
number = 4,
volume = 12,
place = {United States},
year = {Mon Mar 02 00:00:00 EST 2015},
month = {Mon Mar 02 00:00:00 EST 2015}
}

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Cited by: 166 works
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