title: Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors.
abstract: The utility of CRISPR-Cas9 and TALENs for genome editing may be compromised by their off-target activity. We show that integrase-defective lentiviral vectors (IDLVs) can detect such off-target cleavage with a frequency as low as 1%. In the case of Cas9, we find frequent off-target sites with a one-base bulge or up to 13 mismatches between the single guide RNA (sgRNA) and its genomic target, which refines sgRNA design.
authors.affiliation: Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Wang
authors.affiliation: 1] Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA. [2] Bone Marrow Transplantation Center, The First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Yebo
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California, USA.
authors.forename: Xiwei
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California, USA.
authors.forename: Jinhui
authors.initials: J
authors.lastname: Wang
authors.affiliation: 1] Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA. [2] Bone Marrow Transplantation Center, The First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Yingjia
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California, USA.
authors.forename: Zhaojun
authors.initials: Z
authors.lastname: Qiu
authors.affiliation: Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA.
authors.forename: Tammy
authors.initials: T
authors.lastname: Chang
authors.affiliation: Bone Marrow Transplantation Center, The First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: He
authors.initials: H
authors.lastname: Huang
authors.affiliation: Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California, USA.
authors.forename: Ren-Jang
authors.initials: RJ
authors.lastname: Lin
authors.affiliation: Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA.
authors.forename: Jiing-Kuan
authors.initials: JK
authors.lastname: Yee
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D005822:Genetic Vectors; D015894:Genome, Human; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D019426:Integrases / Q000235:genetics; D016086:Lentivirus / Q000201:enzymology / Q000235:genetics*; D017393:RNA Editing / Q000235:genetics*
chemical_list: D019426:Integrases