Full text is available at the source.
Crispr/Cas9‐mediated cleavages facilitate homologous recombination during genetic engineering of a large chromosomal region
CRISPR cuts help precise DNA repair when editing large chromosome regions
AI simplified
Abstract
Homologous recombination efficiency reached up to 16% in mouse embryonic stem cells using Crispr/Cas9 technology.
- An 18.1-kb genomic region around the mTert gene was replaced with a 45.5-kb recombinant fragment.
- A total of 27 mESC clones with heterozygous hmTert/mTert alleles and three clones with homozygous hmTert alleles were obtained.
- The Crispr/Cas9 system induced DNA double-strand breaks, leading to increased homologous recombination efficiency.
- High rates of genomic DNA deletions and mutations were observed at the target sites due to Crispr/Cas9 cleavages.
- This approach may provide a feasible method for manipulating large chromosomal regions in mammalian cells.
AI simplified