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- W2797336915 endingPage "986" @default.
- W2797336915 startingPage "946" @default.
- W2797336915 abstract "CRISPR (clustered regularly interspaced short palindromic repeats) genome-editing experiments offer enormous potential for the evaluation of genomic loci using arrayed single guide RNAs (sgRNAs) or pooled sgRNA libraries. Numerous computational tools are available to help design sgRNAs with optimal on-target efficiency and minimal off-target potential. In addition, computational tools have been developed to analyze deep-sequencing data resulting from genome-editing experiments. However, these tools are typically developed in isolation and oftentimes are not readily translatable into laboratory-based experiments. Here, we present a protocol that describes in detail both the computational and benchtop implementation of an arrayed and/or pooled CRISPR genome-editing experiment. This protocol provides instructions for sgRNA design with CRISPOR (computational tool for the design, evaluation, and cloning of sgRNA sequences), experimental implementation, and analysis of the resulting high-throughput sequencing data with CRISPResso (computational tool for analysis of genome-editing outcomes from deep-sequencing data). This protocol allows for design and execution of arrayed and pooled CRISPR experiments in 4-5 weeks by non-experts, as well as computational data analysis that can be performed in 1-2 d by both computational and noncomputational biologists alike using web-based and/or command-line versions." @default.
- W2797336915 created "2018-04-24" @default.
- W2797336915 creator A5010096006 @default.
- W2797336915 creator A5016855642 @default.
- W2797336915 creator A5038587862 @default.
- W2797336915 creator A5052274720 @default.
- W2797336915 creator A5070184232 @default.
- W2797336915 creator A5071212287 @default.
- W2797336915 creator A5071805086 @default.
- W2797336915 creator A5074055600 @default.
- W2797336915 creator A5078228499 @default.
- W2797336915 creator A5079784932 @default.
- W2797336915 date "2018-04-12" @default.
- W2797336915 modified "2023-10-18" @default.
- W2797336915 title "Integrated design, execution, and analysis of arrayed and pooled CRISPR genome-editing experiments" @default.
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