Inducible and multiplex gene regulation using CRISPR-Cpf1-based transcription factors

Nat Methods. 2017 Dec;14(12):1163-1166. doi: 10.1038/nmeth.4483. Epub 2017 Oct 30.

Abstract

Targeted and inducible regulation of mammalian gene expression is a broadly important capability. We engineered drug-inducible catalytically inactive Cpf1 nuclease fused to transcriptional activation domains to tune the expression of endogenous genes in human cells. Leveraging the multiplex capability of the Cpf1 platform, we demonstrate both synergistic and combinatorial gene expression in human cells. Our work should enable the development of multiplex gene perturbation library screens for understanding complex cellular phenotypes.

MeSH terms

  • Bacterial Proteins / genetics*
  • CRISPR-Cas Systems / genetics*
  • Cell Culture Techniques
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Endonucleases / genetics*
  • Green Fluorescent Proteins / genetics
  • HEK293 Cells
  • Herpes Simplex Virus Protein Vmw65 / genetics
  • Humans
  • Immediate-Early Proteins / genetics
  • Plasmids
  • Recombinant Fusion Proteins / genetics
  • Trans-Activators / genetics
  • Transcription Factor RelA / genetics
  • Transcriptional Activation*
  • Transfection

Substances

  • BRLF1 protein, Human herpesvirus 4
  • Bacterial Proteins
  • Herpes Simplex Virus Protein Vmw65
  • Immediate-Early Proteins
  • Recombinant Fusion Proteins
  • Trans-Activators
  • Transcription Factor RelA
  • Green Fluorescent Proteins
  • Endonucleases