Precise CAG repeat contraction in a Huntington's Disease mouse model is enabled by gene editing with SpCas9-NG

Commun Biol. 2021 Jun 23;4(1):771. doi: 10.1038/s42003-021-02304-w.


The clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 system is a research hotspot in gene therapy. However, the widely used Streptococcus pyogenes Cas9 (WT-SpCas9) requires an NGG protospacer adjacent motif (PAM) for target recognition, thereby restricting targetable disease mutations. To address this issue, we recently reported an engineered SpCas9 nuclease variant (SpCas9-NG) recognizing NGN PAMs. Here, as a feasibility study, we report SpCas9-NG-mediated repair of the abnormally expanded CAG repeat tract in Huntington's disease (HD). By targeting the boundary of CAG repeats with SpCas9-NG, we precisely contracted the repeat tracts in HD-mouse-derived embryonic stem (ES) cells. Further, we confirmed the recovery of phenotypic abnormalities in differentiated neurons and animals produced from repaired ES cells. Our study shows that SpCas9-NG can be a powerful tool for repairing abnormally expanded CAG repeats as well as other disease mutations that are difficult to access with WT-SpCas9.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Associated Protein 9 / physiology*
  • Cell Line
  • Disease Models, Animal
  • Embryonic Stem Cells / cytology
  • Gene Editing*
  • Huntington Disease / genetics*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred ICR
  • Trinucleotide Repeats*


  • CRISPR-Associated Protein 9
  • Cas9 endonuclease Streptococcus pyogenes