Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach

TitleUnraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
Publication TypeJournal Article
Year of Publication2015
AuthorsChari R., Mali P., Moosburner M., Church G.M
JournalNature Methods
Volume12
Pagination823-+
Date Published2015/09
Type of ArticleArticle
ISBN Number1548-7091
Accession NumberWOS:000360586700024
Keywordscas nucleases; cells; cleavage; design; endonuclease; nuclease specificity
Abstract

We developed an in vivo library-on-library methodology to simultaneously assess single guide RNA (sgRNA) activity across similar to 1,400 genomic loci. Assaying across multiple human cell types and end-processing enzymes as well as two Cas9 orthologs, we unraveled underlying nucleotide sequence and epigenetic parameters. Our results and software (http://crispr.med.harvard.edu/sgRNAScorer) enable improved design of reagents, shed light on mechanisms of genome targeting, and provide a generalizable framework to study nucleic acid-nucleic acid interactions and biochemistry in high throughput.

DOI10.1038/nmeth.3473
Short TitleNat. Methods
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