Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Highly-efficient Cas9-mediated transcriptional programming

Alejandro Chavez, Jonathan Scheiman, Suhani Vora, Benjamin W. Pruitt, Marcelle Tuttle, Eswar Iyer, Samira Kiani, Christopher D. Guzman, Daniel J. Wiegand, Dimtry Ter-Ovanesyan, Jonathan L. Braff, Noah Davidsohn, Ron Weiss, John Aach, James J. Collins, George M. Church
doi: https://doi.org/10.1101/012880
Alejandro Chavez
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
2Department of Pathology, Massachusetts General Hospital, Boston, MA 02114, USA
3Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jonathan Scheiman
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
3Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Suhani Vora
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
3Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
4Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Benjamin W. Pruitt
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Marcelle Tuttle
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Eswar Iyer
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
3Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Samira Kiani
4Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
5Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA 02139
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Christopher D. Guzman
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Daniel J. Wiegand
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Dimtry Ter-Ovanesyan
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
3Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jonathan L. Braff
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Noah Davidsohn
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
3Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ron Weiss
4Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
5Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA 02139
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
John Aach
3Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
James J. Collins
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
6Department of Biomedical Engineering and Center of Synthetic Biology, Boston University, Boston, MA 02215, USA
7Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
George M. Church
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
3Department of Genetics, Harvard Medical School, Boston, MA 02115, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

ABSTRACT

The RNA-guided bacterial nuclease Cas9 can be reengineered as a programmable transcription factor by a series of changes to the Cas9 protein in addition to the fusion of a transcriptional activation domain (AD)1–5. However, the modest levels of gene activation achieved by current Cas9 activators have limited their potential applications. Here we describe the development of an improved transcriptional regulator through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to Cas9. We demonstrate its utility in activating expression of endogenous coding and non-coding genes, targeting several genes simultaneously and stimulating neuronal differentiation of induced pluripotent stem cells (iPSCs).

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted December 20, 2014.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Highly-efficient Cas9-mediated transcriptional programming
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Highly-efficient Cas9-mediated transcriptional programming
Alejandro Chavez, Jonathan Scheiman, Suhani Vora, Benjamin W. Pruitt, Marcelle Tuttle, Eswar Iyer, Samira Kiani, Christopher D. Guzman, Daniel J. Wiegand, Dimtry Ter-Ovanesyan, Jonathan L. Braff, Noah Davidsohn, Ron Weiss, John Aach, James J. Collins, George M. Church
bioRxiv 012880; doi: https://doi.org/10.1101/012880
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Highly-efficient Cas9-mediated transcriptional programming
Alejandro Chavez, Jonathan Scheiman, Suhani Vora, Benjamin W. Pruitt, Marcelle Tuttle, Eswar Iyer, Samira Kiani, Christopher D. Guzman, Daniel J. Wiegand, Dimtry Ter-Ovanesyan, Jonathan L. Braff, Noah Davidsohn, Ron Weiss, John Aach, James J. Collins, George M. Church
bioRxiv 012880; doi: https://doi.org/10.1101/012880

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Synthetic Biology
Subject Areas
All Articles
  • Animal Behavior and Cognition (4383)
  • Biochemistry (9599)
  • Bioengineering (7094)
  • Bioinformatics (24865)
  • Biophysics (12615)
  • Cancer Biology (9958)
  • Cell Biology (14354)
  • Clinical Trials (138)
  • Developmental Biology (7950)
  • Ecology (12107)
  • Epidemiology (2067)
  • Evolutionary Biology (15989)
  • Genetics (10926)
  • Genomics (14743)
  • Immunology (9870)
  • Microbiology (23676)
  • Molecular Biology (9485)
  • Neuroscience (50872)
  • Paleontology (369)
  • Pathology (1539)
  • Pharmacology and Toxicology (2683)
  • Physiology (4016)
  • Plant Biology (8657)
  • Scientific Communication and Education (1509)
  • Synthetic Biology (2397)
  • Systems Biology (6436)
  • Zoology (1346)