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Cell Stem Cell DOI:10.1016/j.stem.2014.04.020

Low incidence of off-target mutations in individual CRISPR-Cas9 and TALEN targeted human stem cell clones detected by whole-genome sequencing.

Publication TypeJournal Article
Year of Publication2014
AuthorsVeres, A, Gosis, BS, Ding, Q, Collins, R, Ragavendran, A, Brand, H, Erdin, S, Cowan, CA, Talkowski, ME, Musunuru, K
JournalCell Stem Cell
Volume15
Issue1
Pages27-30
Date Published2014 Jul 03
ISSN1875-9777
KeywordsAdaptor Proteins, Vesicular Transport, Base Sequence, Clone Cells, CRISPR-Cas Systems, Endonucleases, Genetic Engineering, Genome, Humans, Incidence, Molecular Sequence Data, Mutation, Organ Specificity, Pluripotent Stem Cells, Sequence Analysis, DNA
Abstract

Genome editing has attracted wide interest for the generation of cellular models of disease using human pluripotent stem cells and other cell types. CRISPR-Cas systems and TALENs can target desired genomic sites with high efficiency in human cells, but recent publications have led to concern about the extent to which these tools may cause off-target mutagenic effects that could potentially confound disease-modeling studies. Using CRISPR-Cas9 and TALEN targeted human pluripotent stem cell clones, we performed whole-genome sequencing at high coverage in order to assess the degree of mutagenesis across the entire genome. In both types of clones, we found that off-target mutations attributable to the nucleases were very rare. From this analysis, we suggest that, although some cell types may be at risk for off-target mutations, the incidence of such effects in human pluripotent stem cells may be sufficiently low and thus not a significant concern for disease modeling and other applications.

URLhttp://linkinghub.elsevier.com/retrieve/pii/S1934-5909(14)00186-6
DOI10.1016/j.stem.2014.04.020
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/24996167?dopt=Abstract

Alternate JournalCell Stem Cell
PubMed ID24996167
PubMed Central IDPMC4082799
Grant ListU01 HL100408 / HL / NHLBI NIH HHS / United States
R01-HL118744 / HL / NHLBI NIH HHS / United States
R00 HL098364 / HL / NHLBI NIH HHS / United States
R00 MH095867 / MH / NIMH NIH HHS / United States
R00-HL098364 / HL / NHLBI NIH HHS / United States
R01-DK097768 / DK / NIDDK NIH HHS / United States
R00-MH095867 / MH / NIMH NIH HHS / United States
R01 DK097768 / DK / NIDDK NIH HHS / United States
R01 HL118744 / HL / NHLBI NIH HHS / United States