Efficient single-copy HDR by 5' modified long dsDNA donors.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 30156184.
- Also identified by DOI 10.7554/eLife.39468 and PMC identifier 6125127.
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Abstract
CRISPR/Cas9 efficiently induces targeted mutations via non-homologous-end-joining but for genome editing, precise, homology-directed repair (HDR) of endogenous DNA stretches is a prerequisite. To favor HDR, many approaches interfere with the repair machinery or manipulate Cas9 itself. Using Medaka we show that the modification of 5' ends of long dsDNA donors strongly enhances HDR, favors efficient single-copy integration by retaining a monomeric donor conformation thus facilitating successful gene replacement or tagging.
Medical subject headings
- CRISPR-Cas Systems
- DNA
- DNA End-Joining Repair
- Gene Editing
- Recombinational DNA Repair