Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
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New York: Nature Publishing Group US
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English
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New York: Nature Publishing Group US
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Assessing the activity of >1,000 single guide RNAs (sgRNAs) on >1,000 genetic loci with two Cas9 orthologs provides insight into sgRNA design and epigenetic parameters for optimal activity.
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....
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Full title
Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
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TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5292764
Permalink
https://devfeature-collection.sl.nsw.gov.au/record/TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5292764
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ISSN
1548-7091,1548-7105
E-ISSN
1548-7105
DOI
10.1038/nmeth.3473
