Deletion of the GAA repeats from the human frataxin gene using the CRISPR-Cas9 system in YG8R-derived cells and mouse models of Friedreich ataxia

• Published in: Gene therapy Vol. 24; no. 5; pp. 265 - 274
• Main Authors: Ouellet, D L, Cherif, K, Rousseau, J, Tremblay, J P
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.05.2017; Nature Publishing Group
• Subjects: 13/106; 13/109; 13/44; 38/22; 38/23; 38/77; 45/90; 631/1647/1511; 631/1647/1513/1967/3196; 631/208/2489/201; 631/378/1689; 631/61; 64/110; 692/699; 82/80; Analysis; Animal models; Animals; Ataxia; Biomedical and Life Sciences; Biomedicine; Cell Biology; Cell culture; Cell death; Cell Line; Cell lines; Cells, Cultured; CRISPR; CRISPR-Cas Systems; Frataxin; Friedreich Ataxia - genetics; Friedreich Ataxia - therapy; Friedreich's ataxia; Gene deletion; Gene Editing - methods; Gene Expression; Gene Therapy; Genetic aspects; Genetic disorders; Genetic Therapy - methods; Human Genetics; Humans; Iron-Binding Proteins - genetics; Iron-Binding Proteins - metabolism; Mice; Mitochondria; Mutation; Nanotechnology; original-article; Oxidative stress; Protein deficiency; Sequence Deletion; Transcription; Transcription (Genetics); Trinucleotide Repeat Expansion
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2016.89

The Friedreich ataxia is a monogenic disease due to a hyperexpanded GAA triplet located within the first intron of the frataxin gene that causes transcriptional issues. The resulting frataxin protein deficiency leads to a Fe-S cluster biosynthesis dysfunction in the mitochondria and to oxidative stress and cell death. Here we use the CRISPR-Cas9 system to remove the mutated GAA expansion and restore the frataxin gene transcriptional activity and protein level. Both YG8R and YG8sR mouse models and cell lines derived from these mice were used to CRISPR-edited successfully the GAA expansion in vitro and in vivo . Nevertheless, our results suggest the YG8sR as a better and more suitable model for the study of the CRISPR-Cas9 edition of the mutated frataxin gene.