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Title: Precise correction of the dystrophin gene in duchenne muscular dystrophy patient induced pluripotent stem cells by TALEN and CRISPR-Cas9
Authors: Li, Hongmei Lisa
Fujimoto, Naoko
Sasakawa, Noriko
Shirai, Saya
Ohkame, Tokiko
Sakuma, Tetsushi
Tanaka, Michihiro
Amano, Naoki
Watanabe, Akira  kyouindb  KAKEN_id  orcid (unconfirmed)
Sakurai, Hidetoshi  kyouindb  KAKEN_id  orcid (unconfirmed)
Yamamoto, Takashi
Yamanaka, Shinya  kyouindb  KAKEN_id
Hotta, Akitsu  kyouindb  KAKEN_id  orcid (unconfirmed)
Author's alias: 李, 紅梅
笹川, 典子
田中, 道廣
天野, 直己
櫻井, 英俊
山中, 伸弥
堀田, 秋津
Issue Date: 13-Jan-2015
Publisher: Elsevier Inc.
Journal title: Stem Cell Reports
Volume: 4
Issue: 1
Start page: 143
End page: 154
Abstract: Duchenne muscular dystrophy (DMD) is a severe muscle-degenerative disease caused by a mutation in the dystrophin gene. Genetic correction of patient-derived induced pluripotent stem cells (iPSCs) by TALENs or CRISPR-Cas9 holds promise for DMD gene therapy; however, the safety of such nuclease treatment must be determined. Using a unique k-mer database, we systematically identified a unique target region that reduces off-target sites. To restore the dystrophin protein, we performed three correction methods (exon skipping, frameshifting, and exon knockin) in DMD-patient-derived iPSCs, and found that exon knockin was the most effective approach. We further investigated the genomic integrity by karyotyping, copy number variation array, and exome sequencing to identify clones with a minimal mutation load. Finally, we differentiated the corrected iPSCs toward skeletal muscle cells and successfully detected the expression of full-length dystrophin protein. These results provide an important framework for developing iPSC-based gene therapy for genetic disorders using programmable nucleases.
Description: iPS細胞を使った遺伝子修復に成功 --デュシェンヌ型筋ジストロフィーの変異遺伝子を修復--. 京都大学プレスリリース. 2014-11-27.
Rights: © 2015 The Authors. This is an open access article under the CC BY license (
DOI(Published Version): 10.1016/j.stemcr.2014.10.013
PubMed ID: 25434822
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