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

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....

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Detalles Bibliográficos
Autores principales: Chari, Raj, Mali, Prashant, Moosburner, Mark, Church, George M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5292764/
https://www.ncbi.nlm.nih.gov/pubmed/26167643
http://dx.doi.org/10.1038/nmeth.3473
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author Chari, Raj
Mali, Prashant
Moosburner, Mark
Church, George M.
author_facet Chari, Raj
Mali, Prashant
Moosburner, Mark
Church, George M.
author_sort Chari, Raj
collection PubMed
description 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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spelling pubmed-52927642017-02-06 Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach Chari, Raj Mali, Prashant Moosburner, Mark Church, George M. Nat Methods Article 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. 2015-07-13 2015-09 /pmc/articles/PMC5292764/ /pubmed/26167643 http://dx.doi.org/10.1038/nmeth.3473 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Chari, Raj
Mali, Prashant
Moosburner, Mark
Church, George M.
Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
title Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
title_full Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
title_fullStr Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
title_full_unstemmed Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
title_short Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach
title_sort unraveling crispr-cas9 genome engineering parameters via a library-on-library approach
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5292764/
https://www.ncbi.nlm.nih.gov/pubmed/26167643
http://dx.doi.org/10.1038/nmeth.3473
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