Format

Send to

Choose Destination

See 1 citation found by title matching your search:

Nat Methods. 2015 Sep;12(9):823-6. doi: 10.1038/nmeth.3473. Epub 2015 Jul 13.

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

Author information

1
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
2
Department of Bioengineering, University of California, San Diego, La Jolla, California, USA.
3
Scripps Institute of Oceanography, University of California, San Diego, La Jolla, California, USA.
4
Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.

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.

PMID:
26167643
PMCID:
PMC5292764
DOI:
10.1038/nmeth.3473
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center