Controlling transcription in human pluripotent stem cells using CRISPR-effectors

Methods. 2016 May 15;101:36-42. doi: 10.1016/j.ymeth.2015.10.014. Epub 2015 Oct 23.

Abstract

The ability to manipulate transcription in human pluripotent stem cells (hPSCs) is fundamental for the discovery of key genes and mechanisms governing cellular state and differentiation. Recently developed CRISPR-effector systems provide a systematic approach to rapidly test gene function in mammalian cells, including hPSCs. In this review, we discuss recent advances in CRISPR-effector technologies that have been employed to control transcription through gene activation, gene repression, and epigenome engineering. We describe an application of CRISPR-effector mediated transcriptional regulation in hPSCs by targeting a synthetic promoter driving a GFP transgene, demonstrating the ease and effectiveness of CRISPR-effector mediated transcriptional regulation in hPSCs.

Keywords: CRISPR; Cas9; Human pluripotent stem cells; Transcriptional regulation; dCas9-effectors.

Publication types

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

MeSH terms

  • Animals
  • Base Sequence
  • CRISPR-Cas Systems*
  • Cell Culture Techniques
  • HEK293 Cells
  • Humans
  • Pluripotent Stem Cells / physiology*
  • RNA, Guide / genetics
  • Transcription, Genetic*

Substances

  • RNA, Guide