Beditor: A Computational Workflow for Designing Libraries of Guide RNAs for CRISPR-Mediated Base Editing. Issue 2 (1st April 2019)

• Record Type: Journal Article
• Authors: Dandage, Rohan; Després, Philippe C; Yachie, Nozomu; Landry, Christian R
• Is Part Of: Genetics. Volume 212:Issue 2(2019)
• Page Start: 377
• Page End: 385
• Publication Date: 2019-04-01
• Subjects: CRISPR -- base editing -- gene editing -- genome-wide mutagenesis -- gRNA design; Genetics -- Periodicals; 576.5
• DOI: 10.1534/genetics.119.302089
• Languages: English
• ISSNs: 0016-6731

Abstract

Abstract: The biological relevance of the present method pertains to CRISPR-mediated base editing technology that has opened new avenues for scar-free genome-wide mutagenesis and thus to a wide range of applications in genome editing... CRISPR-mediated base editors have opened unique avenues for scar-free genome-wide mutagenesis. Here, we describe a comprehensive computational workflow called beditor that can be broadly adapted for designing guide RNA libraries with a range of CRISPR-mediated base editors, Protospacer Adjacent Motif (PAM) recognition sequences, and genomes of many species. Additionally, to assist users in selecting the best sets of guide RNAs for their experiments, a priori estimates of editing efficiency, called beditor scores, are calculated. These beditor scores are intended to select guide RNAs that conform to requirements for optimal base editing: the editable base falls within maximum activity window of the CRISPR-mediated base editor and produces nonconfounding mutational effects with minimal predicted off-target effects. We demonstrate the utility of the software by designing guide RNAs for base editing to model or correct thousands of clinically important human disease mutations.