Correction of a genetic disease in mouse via use of CRISPR-Cas9

Cell Stem Cell. 2013 Dec 5;13(6):659-62. doi: 10.1016/j.stem.2013.10.016.

Abstract

The CRISPR-Cas9 system has been employed to generate mutant alleles in a range of different organisms. However, so far there have not been reports of use of this system for efficient correction of a genetic disease. Here we show that mice with a dominant mutation in Crygc gene that causes cataracts could be rescued by coinjection into zygotes of Cas9 mRNA and a single-guide RNA (sgRNA) targeting the mutant allele. Correction occurred via homology-directed repair (HDR) based on an exogenously supplied oligonucleotide or the endogenous WT allele, with only rare evidence of off-target modifications. The resulting mice were fertile and able to transmit the corrected allele to their progeny. Thus, our study provides proof of principle for use of the CRISPR-Cas9 system to correct genetic disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • CRISPR-Associated Proteins / metabolism*
  • Cataract / genetics*
  • Cataract / therapy*
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Disease Models, Animal
  • Genetic Therapy*
  • Mice
  • Molecular Sequence Data
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • gamma-Crystallins / genetics
  • gamma-Crystallins / therapeutic use

Substances

  • CRISPR-Associated Proteins
  • RNA, Messenger
  • gamma-Crystallins