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発表内容

Title:
CRISPR system for genome editing gene therapy in Duchenne muscular dystrophy iPS cells

 

Akitsu Hotta, Ph.D.
Center for iPS Cell Reserach and Application (CiRA), Department of Life Science Frontiers, Kyoto University

 

Abstract:
Duchenne Muscular Dystrophy (DMD) is one of the most common and severe inherited neuromuscular disease caused by the loss-of-function mutations in Dystrophin gene on X chromosome. Currently, no effective treatment available for DMD, including gene augmentation therapy, since large size of the Dystrophin gene hamper the delivery by viral vectors. Exon skipping to modulate mRNA splicing patterns using antisense oligonucleotide is a promising approach currently investigated in clinical trials, however, the effect of antisense oligos is transient. To overcome those issues, gene correction approach is eagerly desired1).
Recent progress on targeted gene editing by CRISPR-Cas9 system have evolutionally broaden our ability to precisely modify the genomic sequence at desired locus in iPS cells2, 3). In fact, we recently demonstrated that CRISPR-Cas9 is a powerful system to correct out-of-frame mutation of the Dystrophin gene in DMD patient-derived iPS cells4). At the seminar, I will summerize our recent efforts to enhance genome editing efficiency in iPS cells5), and to reduce off-target mutagenesis6), since gene-correction by CRISPR system holds a promise to serve as a novel genome editing gene therapy for DMD.

 

References:
1) Hotta A.: “Genome editing gene therapy for Duchenne muscular dystrophy.”
J. Neuromuscul. Dis., 2015 Sep 22, 2 (4), 343-355

2) Hotta A, and Yamanaka S.: “From genomics to gene therapy: induced pluripotent stem cells meet genome editing.” Annu. Rev. Genet., 2015 Sep 25.

3) Li HL, Nakano T, Hotta A.: “Genetic correction using engineered nucleases for gene therapy applications.” Development Growth & Differentiation, 2014; Vol.56 (1): p63-77.

4) Li HL, Fujimoto N, Sasakawa N, Shirai S, Ohkame T, Sakuma T, Tanaka M, Amano N, Watanabe A, Sakurai H, Yamamoto T, Yamanaka S, and Hotta A.: “Precise correction of the dystrophin gene in Duchenne muscular dystrophy patient induced pluripotent stem cells by TALEN and CRISPR-Cas9.” Stem Cell Reports, 2015; Vol.4 (1): p143-154.

5) Li HL, Gee P, Ishida K, and Hotta A.: “Efficient genomic correction methods in human iPS cells using CRISPR-Cas9 system.” Methods, 2015 Oct 23, pii: S1046-2023(15)30133-X.

6) Ishida K, Gee P, and Hotta A.: “Minimizing off-target mutagenesis risks caused by programmable nucleases.” Int. J. Mol. Sci. 2015 Oct 16, 16 (10), 24751-24771