PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 26644285.
- Also identified by DOI 10.1038/ncomms10124 and PMC identifier 4686818.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
In CRISPR-Cas9 genome editing, the underlying principles for selecting guide RNA (gRNA) sequences that would ensure for efficient target site modification remain poorly understood. Here we show that target sites harbouring multiple protospacer adjacent motifs (PAMs) are refractory to Cas9-mediated repair in situ. Thus we refine which substrates should be avoided in gRNA design, implicating PAM density as a novel sequence-specific feature that inhibits in vivo Cas9-driven DNA modification.
Medical subject headings
- CRISPR-Cas Systems
- DNA Cleavage
- Nucleotide Motifs
- RNA Editing
- RNA, Guide, CRISPR-Cas Systems