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Cell Stem Cell. 2014 Jul 3;15(1):31-6. doi: 10.1016/j.stem.2014.06.016.

Targeted gene correction minimally impacts whole-genome mutational load in human-disease-specific induced pluripotent stem cell clones.

Author information

1
Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA.
2
BGI, Beishan Industrial Zone, Yantian District, Shenzhen 518083, China.
3
Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
4
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
5
BGI, Beishan Industrial Zone, Yantian District, Shenzhen 518083, China; Institute of Digestive Disease and the Department of Medicine and Therapeutics, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Hong Kong.
6
BGI, Beishan Industrial Zone, Yantian District, Shenzhen 518083, China; Department of Computational Medicine and Bioinformatics, Medical School, University of Michigan, 500 South State Street, Ann Arbor, MI 48109, USA.
7
Integrative Genomics and Bioinformatics Core, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA.
8
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; Center for Age-related Diseases (CAD), Beijing, China; Beijing Institute for Brain Disorders, Beijing 100069, China. Electronic address: ghliu@ibp.ac.cn.
9
BGI, Beishan Industrial Zone, Yantian District, Shenzhen 518083, China. Electronic address: liyr@genomics.cn.
10
Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA. Electronic address: belmonte@salk.edu.

Abstract

The utility of genome editing technologies for disease modeling and developing cellular therapies has been extensively documented, but the impact of these technologies on mutational load at the whole-genome level remains unclear. We performed whole-genome sequencing to evaluate the mutational load at single-base resolution in individual gene-corrected human induced pluripotent stem cell (hiPSC) clones in three different disease models. In single-cell clones, gene correction by helper-dependent adenoviral vector (HDAdV) or Transcription Activator-Like Effector Nuclease (TALEN) exhibited few off-target effects and a low level of sequence variation, comparable to that accumulated in routine hiPSC culture. The sequence variants were randomly distributed and unique to individual clones. We also combined both technologies and developed a TALEN-HDAdV hybrid vector, which significantly increased gene-correction efficiency in hiPSCs. Therefore, with careful monitoring via whole-genome sequencing it is possible to apply genome editing to human pluripotent cells with minimal impact on genomic mutational load.

PMID:
24996168
PMCID:
PMC4144407
DOI:
10.1016/j.stem.2014.06.016
[Indexed for MEDLINE]
Free PMC Article

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