Augmenting and directing long-range CRISPR-mediated activation in human cells

Nat Methods. 2021 Sep;18(9):1075-1081. doi: 10.1038/s41592-021-01224-1. Epub 2021 Aug 5.


Epigenetic editing is an emerging technology that uses artificial transcription factors (aTFs) to regulate expression of a target gene. Although human genes can be robustly upregulated by targeting aTFs to promoters, the activation induced by directing aTFs to distal transcriptional enhancers is substantially less robust and consistent. Here we show that long-range activation using CRISPR-based aTFs in human cells can be made more efficient and reliable by concurrently targeting an aTF to the target gene promoter. We used this strategy to direct target gene choice for enhancers capable of regulating more than one promoter and to achieve allele-selective activation of human genes by targeting aTFs to single-nucleotide polymorphisms embedded in distally located sequences. Our results broaden the potential applications of the epigenetic editing toolbox for research and therapeutics.

Publication types

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

MeSH terms

  • Alleles
  • Apolipoprotein C-III / genetics
  • Apolipoproteins A / genetics
  • Cell Line
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Enhancer Elements, Genetic
  • Gene Targeting / methods*
  • Humans
  • Interleukin-2 Receptor alpha Subunit / genetics
  • MyoD Protein / genetics
  • Polymorphism, Single Nucleotide
  • Promoter Regions, Genetic*
  • Transcription Factors / genetics*
  • Transcriptional Activation
  • beta-Globins / genetics


  • APOC3 protein, human
  • Apolipoprotein C-III
  • Apolipoproteins A
  • IL2RA protein, human
  • Interleukin-2 Receptor alpha Subunit
  • MyoD Protein
  • MyoD1 myogenic differentiation protein
  • Transcription Factors
  • apolipoprotein A-IV
  • beta-Globins