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

ABSTRACT: We developed an in vivo library-on-library methodology to simultaneously assess single guide RNA (sgRNA) activity across ?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.

SUBMITTER: Chari R

PROVIDER: S-EPMC5292764 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature

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

Chari Raj R Mali Prashant P Moosburner Mark M Church George M GM

Nature methods 20150713 9

We developed an in vivo library-on-library methodology to simultaneously assess single guide RNA (sgRNA) activity across ∼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 framewor ...[more]

PMID: 26167643

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