A G542X cystic fibrosis mouse model for examining nonsense mutation directed therapies

PLoS One. 2018 Jun 20;13(6):e0199573. doi: 10.1371/journal.pone.0199573. eCollection 2018.

Abstract

Nonsense mutations are present in 10% of patients with CF, produce a premature termination codon in CFTR mRNA causing early termination of translation, and lead to lack of CFTR function. There are no currently available animal models which contain a nonsense mutation in the endogenous Cftr locus that can be utilized to test nonsense mutation therapies. In this study, we create a CF mouse model carrying the G542X nonsense mutation in Cftr using CRISPR/Cas9 gene editing. The G542X mouse model has reduced Cftr mRNA levels, demonstrates absence of CFTR function, and displays characteristic manifestations of CF mice such as reduced growth and intestinal obstruction. Importantly, CFTR restoration is observed in G542X intestinal organoids treated with G418, an aminoglycoside with translational readthrough capabilities. The G542X mouse model provides an invaluable resource for the identification of potential therapies of CF nonsense mutations as well as the assessment of in vivo effectiveness of these potential therapies targeting nonsense mutations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems
  • Cells, Cultured
  • Codon, Nonsense*
  • Cystic Fibrosis / genetics
  • Cystic Fibrosis / metabolism
  • Cystic Fibrosis / therapy*
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
  • Disease Models, Animal*
  • Female
  • Genetic Therapy / methods*
  • Intestines
  • Male
  • Mice, Inbred C57BL
  • Mice, Transgenic*
  • Organoids / drug effects
  • Organoids / metabolism
  • RNA, Messenger / metabolism
  • Tissue Culture Techniques

Substances

  • Cftr protein, mouse
  • Codon, Nonsense
  • RNA, Messenger
  • Cystic Fibrosis Transmembrane Conductance Regulator

Grant support

This work was supported by grants from the Cystic Fibrosis Foundation Therapeutics (Hodges15XX0 to CAH and Conlon15XX0 to RAC, https://www.cff.org/ CFF). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.