title: Low incidence of off-target mutations in individual CRISPR-Cas9 and TALEN targeted human stem cell clones detected by whole-genome sequencing.
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.
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Adrian
authors.initials: A
authors.lastname: Veres
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Bridget S
authors.initials: BS
authors.lastname: Gosis
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Qiurong
authors.initials: Q
authors.lastname: Ding
authors.affiliation: Molecular Neurogenetics Unit, Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, MA 02114, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Collins
authors.affiliation: Molecular Neurogenetics Unit, Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, MA 02114, USA.
authors.forename: Ashok
authors.initials: A
authors.lastname: Ragavendran
authors.affiliation: Molecular Neurogenetics Unit, Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, MA 02114, USA.
authors.forename: Harrison
authors.initials: H
authors.lastname: Brand
authors.affiliation: Molecular Neurogenetics Unit, Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, MA 02114, USA.
authors.forename: Serkan
authors.initials: S
authors.lastname: Erdin
authors.forename: Chad A
authors.initials: CA
authors.lastname: Cowan
authors.affiliation: Molecular Neurogenetics Unit, Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Neurology, Harvard Medical School, Boston, MA 02115, USA; Broad Institute, Cambridge, MA 02142, USA.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Talkowski
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Department of Medicine, Harvard Medical School, Boston, MA 02115, USA; Broad Institute, Cambridge, MA 02142, USA; Division of Cardiovascular Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA. Electronic address: kiranmusunuru@gmail.com.
authors.forename: Kiran
authors.initials: K
authors.lastname: Musunuru
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics; D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002999:Clone Cells; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering; D016678:Genome / Q000235:genetics; D006801:Humans; D015994:Incidence; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009928:Organ Specificity; D039904:Pluripotent Stem Cells / Q000502:physiology*; D017422:Sequence Analysis, DNA / Q000379:methods*; D000099183:Sortilin
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D004720:Endonucleases; D000099183:Sortilin