Highly efficient Cas9-mediated transcriptional programming

Alejandro Chavez; Jonathan Scheiman; Suhani Vora; Benjamin W Pruitt; Marcelle Tuttle; Eswar P R Iyer; Shuailiang Lin; Samira Kiani; Christopher D Guzman; Daniel J Wiegand; Dmitry Ter-Ovanesyan; Jonathan L Braff; Noah Davidsohn; Benjamin E Housden; Norbert Perrimon; Ron Weiss; John Aach; James J Collins; George M Church

doi:10.1038/nmeth.3312

Highly efficient Cas9-mediated transcriptional programming

Nat Methods. 2015 Apr;12(4):326-8. doi: 10.1038/nmeth.3312. Epub 2015 Mar 2.

Authors

Alejandro Chavez¹, Jonathan Scheiman², Suhani Vora³, Benjamin W Pruitt⁴, Marcelle Tuttle⁴, Eswar P R Iyer², Shuailiang Lin⁵, Samira Kiani⁶, Christopher D Guzman⁴, Daniel J Wiegand⁴, Dmitry Ter-Ovanesyan², Jonathan L Braff⁴, Noah Davidsohn², Benjamin E Housden⁷, Norbert Perrimon⁷, Ron Weiss⁸, John Aach⁹, James J Collins¹⁰, George M Church²

Affiliations

¹ 1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA. [3] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
² 1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
³ 1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. [3] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
⁴ Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.
⁵ 1] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA. [3] School of Life Sciences, Tsinghua University, Beijing, China.
⁶ 1] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
⁷ 1] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.
⁸ 1] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [3] Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
⁹ Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
¹⁰ 1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA. [2] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [3] Institute for Medical Engineering &Science, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [4] Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.

Abstract

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

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell Differentiation / genetics
Endonucleases* / genetics
Genetic Techniques*
HEK293 Cells
Humans
Induced Pluripotent Stem Cells
Neurons / cytology
RNA, Guide, CRISPR-Cas Systems
Staphylococcus aureus
Transcriptional Activation*

Substances

Endonucleases

Abstract

Publication types

MeSH terms

Substances

Grants and funding