Publication: Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
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2017
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Chari, Raj, Prashant Mali, Mark Moosburner, and George M. Church. 2017. “Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach.” Nature methods 12 (9): 823-826. doi:10.1038/nmeth.3473. http://dx.doi.org/10.1038/nmeth.3473.
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Abstract
We develop 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, end-processing enzymes, and two Cas9 orthologs, we unravel underlying nucleotide sequence and epigenetic parameters. Our results 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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