論文 - 詳細
| 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 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 46 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 3.0 |
| リソース情報 | |
| ヒト・動物細胞 | 409B2(HPS0076) |