In Vivo Target Gene Activation via CRISPR/Cas9-Mediated Trans-epigenetic Modulation

Cell. 2017 Dec 14;171(7):1495-1507.e15. doi: 10.1016/j.cell.2017.10.025. Epub 2017 Dec 7.


Current genome-editing systems generally rely on inducing DNA double-strand breaks (DSBs). This may limit their utility in clinical therapies, as unwanted mutations caused by DSBs can have deleterious effects. CRISPR/Cas9 system has recently been repurposed to enable target gene activation, allowing regulation of endogenous gene expression without creating DSBs. However, in vivo implementation of this gain-of-function system has proven difficult. Here, we report a robust system for in vivo activation of endogenous target genes through trans-epigenetic remodeling. The system relies on recruitment of Cas9 and transcriptional activation complexes to target loci by modified single guide RNAs. As proof-of-concept, we used this technology to treat mouse models of diabetes, muscular dystrophy, and acute kidney disease. Results demonstrate that CRISPR/Cas9-mediated target gene activation can be achieved in vivo, leading to measurable phenotypes and amelioration of disease symptoms. This establishes new avenues for developing targeted epigenetic therapies against human diseases. VIDEO ABSTRACT.

Keywords: CRISPR/Cas9; chromatin remodeling; disease model; epigenetic modification; epigenetic therapy; gene editing; muscular dystrophy; regenerative medicine; stem cells; transdifferentiation.

MeSH terms

  • Animals
  • Base Sequence
  • CRISPR-Cas Systems*
  • Disease Models, Animal
  • Dystrophin / genetics
  • Epigenesis, Genetic*
  • Gene Targeting / methods*
  • Genetic Therapy / methods*
  • Interleukin-10 / genetics
  • Klotho Proteins
  • Membrane Proteins / genetics
  • Mice
  • Mice, Inbred C57BL
  • Muscular Dystrophy, Duchenne / genetics*
  • Muscular Dystrophy, Duchenne / therapy*
  • Transcriptional Activation
  • Utrophin / genetics*


  • Dystrophin
  • Klb protein, mouse
  • Membrane Proteins
  • Utrophin
  • apo-dystrophin 1
  • Interleukin-10
  • Klotho Proteins