論文 - 詳細
| RRC ID | 55956 |
|---|---|
| 著者 | Kawabe Y, Komatsu S, Komatsu S, Murakami M, Ito A, Sakuma T, Nakamura T, Yamamoto T, Kamihira M. |
| タイトル | Targeted knock-in of an scFv-Fc antibody gene into the hprt locus of Chinese hamster ovary cells using CRISPR/Cas9 and CRIS-PITCh systems. |
| ジャーナル | J Biosci Bioeng |
| Abstract |
Chinese hamster ovary (CHO) cells have been used as host cells for the production of pharmaceutical proteins. For the high and stable production of target proteins, the transgene should be integrated into a suitable genomic locus of host cells. Here, we generated knock-in CHO cells, in which transgene cassettes without a vector backbone sequence were integrated into the hprt locus of the CHO genome using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and CRISPR-mediated precise integration into target chromosome (CRIS-PITCh) systems. We investigated the efficiency of targeted knock-in of transgenes using these systems. As a practical example, we generated knock-in CHO cells producing an scFv-Fc antibody using the CRIS-PITCh system mediated by microhomology sequences for targeting. We found that the CRIS-PITCh system can facilitate targeted knock-in for CHO cell engineering. |
| 巻・号 | 125(5) |
| ページ | 599-605 |
| 公開日 | 2018-5-1 |
| DOI | 10.1016/j.jbiosc.2017.12.003 |
| PII | S1389-1723(17)31069-1 |
| PMID | 29295784 |
| MeSH | Animals CHO Cells CRISPR-Cas Systems* Cricetinae Cricetulus Gene Knock-In Techniques / methods* Genes, Immunoglobulin Genetic Loci Hypoxanthine Phosphoribosyltransferase / genetics* Immunoglobulin Fc Fragments / genetics* Mutagenesis, Insertional / methods* Single-Chain Antibodies / genetics* Transgenes |
| IF | 2.032 |
| 引用数 | 9 |
| 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 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 13 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | CHO-K1 |