RRC ID 58000
著者 Riesenberg S, Chintalapati M, Macak D, Kanis P, Maricic T, Pääbo S.
タイトル Simultaneous precise editing of multiple genes in human cells.
ジャーナル Nucleic Acids Res
Abstract When double-strand breaks are introduced in a genome by CRISPR they are repaired either by non-homologous end joining (NHEJ), which often results in insertions or deletions (indels), or by homology-directed repair (HDR), which allows precise nucleotide substitutions to be introduced if a donor oligonucleotide is provided. Because NHEJ is more efficient than HDR, the frequency with which precise genome editing can be achieved is so low that simultaneous editing of more than one gene has hitherto not been possible. Here, we introduced a mutation in the human PRKDC gene that eliminates the kinase activity of the DNA-dependent protein kinase catalytic subunit (DNA-PKcs). This results in an increase in HDR irrespective of cell type and CRISPR enzyme used, sometimes allowing 87% of chromosomes in a population of cells to be precisely edited. It also allows for precise editing of up to four genes simultaneously (8 chromosomes) in the same cell. Transient inhibition of DNA-PKcs by the kinase inhibitor M3814 is similarly able to enhance precise genome editing.
巻・号 47(19)
ページ e116
公開日 2019-11-4
DOI 10.1093/nar/gkz669
PII 5545003
PMID 31392986
PMC PMC6821318
MeSH CRISPR-Cas Systems / genetics Chromosomes DNA Breaks, Double-Stranded* DNA End-Joining Repair / genetics DNA-Activated Protein Kinase / genetics* Gene Editing / methods* HEK293 Cells Humans INDEL Mutation / genetics Oligonucleotides / genetics Recombinational DNA Repair / genetics* Sequence Deletion / genetics
IF 11.502
引用数 4
リソース情報
ヒト・動物細胞 409B2(HPS0076)