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Hum Mol Genet. 2015 Jul 1;24(13):3764-74. doi: 10.1093/hmg/ddv120. Epub 2015 Apr 9.

Functional disruption of the dystrophin gene in rhesus monkey using CRISPR/Cas9.

Author information

1
Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China, Kunming Biomed International and National Engineering Research Center of Biomedicine and Animal Science, Kunming, 650500, China and.
2
State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 10010, China.
3
Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China, Kunming Biomed International and National Engineering Research Center of Biomedicine and Animal Science, Kunming, 650500, China and.
4
Department of Physiology, College of Medicine, National Cheng Kung University, Tainan 70101, Taiwan.
5
Department of Human Genetics, Emory University School of Medicine, Atlanta, Georgia, 30322, USA.
6
Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China, Kunming Biomed International and National Engineering Research Center of Biomedicine and Animal Science, Kunming, 650500, China and xli2@emory.edu wji@kbimed.com.
7
State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 10010, China, Department of Human Genetics, Emory University School of Medicine, Atlanta, Georgia, 30322, USA, xli2@emory.edu wji@kbimed.com.

Abstract

CRISPR/Cas9 has been used to genetically modify genomes in a variety of species, including non-human primates. Unfortunately, this new technology does cause mosaic mutations, and we do not yet know whether such mutations can functionally disrupt the targeted gene or cause the pathology seen in human disease. Addressing these issues is necessary if we are to generate large animal models of human diseases using CRISPR/Cas9. Here we used CRISPR/Cas9 to target the monkey dystrophin gene to create mutations that lead to Duchenne muscular dystrophy (DMD), a recessive X-linked form of muscular dystrophy. Examination of the relative targeting rate revealed that Crispr/Cas9 targeting could lead to mosaic mutations in up to 87% of the dystrophin alleles in monkey muscle. Moreover, CRISPR/Cas9 induced mutations in both male and female monkeys, with the markedly depleted dystrophin and muscle degeneration seen in early DMD. Our findings indicate that CRISPR/Cas9 can efficiently generate monkey models of human diseases, regardless of inheritance patterns. The presence of degenerated muscle cells in newborn Cas9-targeted monkeys suggests that therapeutic interventions at the early disease stage may be effective at alleviating the myopathy.

PMID:
25859012
PMCID:
PMC5007610
DOI:
10.1093/hmg/ddv120
[Indexed for MEDLINE]
Free PMC Article

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