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CasKAS: direct profiling of genome-wide dCas9 and Cas9 specificity using ssDNA mapping.
Marinov, Georgi K; Kim, Samuel H; Bagdatli, S Tansu; Higashino, Soon Il; Trevino, Alexandro E; Tycko, Josh; Wu, Tong; Bintu, Lacramioara; Bassik, Michael C; He, Chuan; Kundaje, Anshul; Greenleaf, William J.
Afiliación
  • Marinov GK; Department of Genetics, School of Medicine, Stanford University, Stanford, CA, 94305, USA. GKM359@gmail.com.
  • Kim SH; Cancer Biology Program, School of Medicine, Stanford University, Stanford, CA, 94305, USA.
  • Bagdatli ST; Department of Genetics, School of Medicine, Stanford University, Stanford, CA, 94305, USA.
  • Higashino SI; Department of Genetics, School of Medicine, Stanford University, Stanford, CA, 94305, USA.
  • Trevino AE; Center for Personal Dynamic Regulomes, Stanford University, Stanford, CA, 94305, USA.
  • Tycko J; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.
  • Wu T; Department of Genetics, School of Medicine, Stanford University, Stanford, CA, 94305, USA.
  • Bintu L; Department of Chemistry, The University of Chicago, Chicago, IL, 60637, USA.
  • Bassik MC; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.
  • He C; Department of Genetics, School of Medicine, Stanford University, Stanford, CA, 94305, USA.
  • Kundaje A; Chemistry, Engineering, and Medicine for Human Health (ChEM-H), Stanford University, Stanford, CA, 94305, USA.
  • Greenleaf WJ; Department of Chemistry, The University of Chicago, Chicago, IL, 60637, USA.
Genome Biol ; 24(1): 85, 2023 04 21.
Article en En | MEDLINE | ID: mdl-37085898
Detecting and mitigating off-target activity is critical to the practical application of CRISPR-mediated genome and epigenome editing. While numerous methods have been developed to map Cas9 binding specificity genome-wide, they are generally time-consuming and/or expensive, and not applicable to catalytically dead CRISPR enzymes. We have developed CasKAS, a rapid, inexpensive, and facile assay for identifying off-target CRISPR enzyme binding and cleavage by chemically mapping the unwound single-stranded DNA structures formed upon binding of a sgRNA-loaded Cas9 protein. We demonstrate this method in both in vitro and in vivo contexts.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ADN de Cadena Simple / Sistemas CRISPR-Cas Idioma: En Revista: Genome Biol Asunto de la revista: BIOLOGIA MOLECULAR / GENETICA Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
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PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ADN de Cadena Simple / Sistemas CRISPR-Cas Idioma: En Revista: Genome Biol Asunto de la revista: BIOLOGIA MOLECULAR / GENETICA Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos