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Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors.
Wang, Xiaoling; Wang, Yebo; Wu, Xiwei; Wang, Jinhui; Wang, Yingjia; Qiu, Zhaojun; Chang, Tammy; Huang, He; Lin, Ren-Jang; Yee, Jiing-Kuan.
Afiliación
  • Wang X; Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA.
  • Wang Y; 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.
  • Wu X; Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California, USA.
  • Wang J; Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California, USA.
  • Wang Y; 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.
  • Qiu Z; Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California, USA.
  • Chang T; Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA.
  • Huang H; Bone Marrow Transplantation Center, The First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, China.
  • Lin RJ; Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, California, USA.
  • Yee JK; Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA.
Nat Biotechnol ; 33(2): 175-8, 2015 Feb.
Article en En | MEDLINE | ID: mdl-25599175
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Edición de ARN / Lentivirus / Sistemas CRISPR-Cas Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Nat Biotechnol Asunto de la revista: BIOTECNOLOGIA Año: 2015 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Edición de ARN / Lentivirus / Sistemas CRISPR-Cas Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Nat Biotechnol Asunto de la revista: BIOTECNOLOGIA Año: 2015 Tipo del documento: Article País de afiliación: Estados Unidos