論文 - 詳細
| RRC ID | 33118 |
|---|---|
| 著者 | Nakade S, Tsubota T, Sakane Y, Kume S, Sakamoto N, Obara M, Daimon T, Sezutsu H, Yamamoto T, Sakuma T, Suzuki KT. |
| タイトル | Microhomology-mediated end-joining-dependent integration of donor DNA in cells and animals using TALENs and CRISPR/Cas9. |
| ジャーナル | Nat Commun |
| Abstract |
Genome engineering using programmable nucleases enables homologous recombination (HR)-mediated gene knock-in. However, the labour used to construct targeting vectors containing homology arms and difficulties in inducing HR in some cell type and organisms represent technical hurdles for the application of HR-mediated knock-in technology. Here, we introduce an alternative strategy for gene knock-in using transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) mediated by microhomology-mediated end-joining, termed the PITCh (Precise Integration into Target Chromosome) system. TALEN-mediated PITCh, termed TAL-PITCh, enables efficient integration of exogenous donor DNA in human cells and animals, including silkworms and frogs. We further demonstrate that CRISPR/Cas9-mediated PITCh, termed CRIS-PITCh, can be applied in human cells without carrying the plasmid backbone sequence. Thus, our PITCh-ing strategies will be useful for a variety of applications, not only in cultured cells, but also in various organisms, including invertebrates and vertebrates. |
| 巻・号 | 5 |
| ページ | 5560 |
| 公開日 | 2014-11-20 |
| DOI | 10.1038/ncomms6560 |
| PII | ncomms6560 |
| PMID | 25410609 |
| PMC | PMC4263139 |
| MeSH | Animals Base Sequence Bombyx CRISPR-Cas Systems* DNA / metabolism* Deoxyribonucleases* Gene Knock-In Techniques / methods* Genetic Engineering Genetic Vectors Homologous Recombination Humans Molecular Sequence Data Plasmids Saccharomyces cerevisiae Xenopus |
| IF | 12.121 |
| 引用数 | 214 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 137 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 21.0 |
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