Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 26167643.
- Also identified by DOI 10.1038/nmeth.3473 and PMC identifier 5292764.
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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.
Medical subject headings
- Chromosome Mapping
- Clustered Regularly Interspaced Short Palindromic Repeats
- Genetic Engineering
- Genome, Human
- Genomic Library
- RNA