RT Journal Article
SR Electronic
T1 CRISPR-Cas9 targeted deletion of the <em>C9orf72</em> repeat expansion mutation corrects cellular phenotypes in patient-derived iPS cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 051193
DO 10.1101/051193
A1 Mochtar Pribadi
A1 Zhongan Yang
A1 Tanya S. Kim
A1 Elliot W. Swartz
A1 Alden Y. Huang
A1 Jason A. Chen
A1 Deepika Dokuru
A1 Jaeyun Baek
A1 Fuying Gao
A1 Andrea T. Fua
A1 Kevin Wojta
A1 Qing Wang
A1 Anna Karydas
A1 Jamie Fong
A1 Ed Lezcano
A1 Stephanie Ng
A1 Farid F. Chehab
A1 Harry V. Vinters
A1 Bruce L. Miller
A1 Giovanni Coppola
YR 2016
UL http://biorxiv.org/content/early/2016/05/02/051193.abstract
AB The large hexanucleotide (GGGGCC) repeat expansion in the non-coding promoter region of C9orf72 is the leading cause of Frontotemporal Dementia (FTD) and Amyotrophic Lateral Sclerosis (ALS). Mechanisms underlying neurodegeneration are not clear, and both a C9orf72 loss of function and a gain of toxicity, in the form of RNA foci or dipeptide repeat deposition, are implicated. CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9-mediated genome editing is an attractive strategy for disease modeling and therapeutic intervention. Here we show that this system can be utilized to completely remove the large repeat expansion mutation within C9orf72 in patient-derived induced pluripotent stem (iPS) cells. Removal of the mutation prevented RNA foci formation and promoter hypermethylation, two phenotypes of the C9orf72 mutation. Interestingly, these changes did not significantly alter C9orf72 expression at the mRNA or protein level. This work provides a proof-of-principle for the use of CRISPR-Cas9-mediated excision of the pathogenic C9orf72 repeat expansion as a therapeutic strategy in FTD/ALS.One Sentence Summary CRISPR-Cas9-mediated excision of the large C9orf72 repeat expansion mutation rescues RNA foci formation and promoter hypermethylation without altering C9orf72 transcript and protein expression.