Highly efficient Cas9-mediated transcriptional programming

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

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*
  • Staphylococcus aureus
  • Transcriptional Activation*

Substances

  • RNA, Guide
  • Endonucleases