CRISPR/Cas9-mediated targeted gene correction in amyotrophic lateral sclerosis patient iPSCs
Lixia Wang1,2,3, Fei Yi4, Lina Fu1,3, Jiping Yang1,3, Si Wang1,3, Zhaoxia Wang5, Keiichiro Suzuki6,7, Liang Sun9, Xiuling Xu1, Yang Yu8, Jie Qiao8, Juan Carlos Izpisua Belmonte6, Ze Yang9, Yun Yuan5(), Jing Qu2,3(), Guang-Hui Liu1,3,10,11()
1. National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China 2. State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China 3. University of Chinese Academy of Sciences, Beijing 100049, China 4. Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA 5. Department of Neurology, Peking University First Hospital, Beijing 100034, China 6. Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA 7. Universidad Católica San Antonio de Murcia (UCAM), Campus de los Jerónimos, N 135 Guadalupe, 30107 Murcia, Spain 8. Department of Gynecology and Obstetrics, Peking University Third Hospital, Beijing 100191, China 9. Beijing Hospital of the Ministry of Health, Beijing 100730, China 10. Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou 510632, China 11. Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China
Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease with cellular and molecular mechanisms yet to be fully described. Mutations in a number of genes including SOD1 and FUS are associated with familial ALS. Here we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts of familial ALS patients bearing SOD1+/A272C and FUS+/G1566A mutations, respectively. We further generated gene corrected ALS iPSCs using CRISPR/Cas9 system. Genome-wide RNA sequencing (RNA-seq) analysis ofmotor neurons derived from SOD1+/A272C and corrected iPSCs revealed 899 aberrant transcripts. Our work may shed light on discovery of early biomarkers and pathways dysregulated in ALS, as well as provide a basis for novel therapeutic strategies to treat ALS.
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