Highly efficient Cas9-mediated transcriptional programming

  • Nat Methods. 2015 Apr;12(4):326-8. doi: 10.1038/nmeth.3312.
Alejandro Chavez  1 ,  Jonathan Scheiman  2 ,  Suhani Vora  3 ,  Benjamin W Pruitt  4 ,  Marcelle Tuttle  4 ,  Eswar P R Iyer  2 ,  Shuailiang Lin  5 ,  Samira Kiani  6 ,  Christopher D Guzman  4 ,  Daniel J Wiegand  4 ,  Dmitry Ter-Ovanesyan  2 ,  Jonathan L Braff  4 ,  Noah Davidsohn  2 ,  Benjamin E Housden  7 ,  Norbert Perrimon  7 ,  Ron Weiss  8 ,  John Aach  9 ,  James J Collins  10 ,  George M Church  2
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.
PMID: 25730490 DOI: 10.1038/nmeth.3312
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).