論文 - 詳細
| RRC ID | 57447 |
|---|---|
| 著者 | Honda A, Tachibana R, Hamada K, Morita K, Mizuno N, Morita K, Asano M. |
| タイトル | Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization. |
| ジャーナル | Sci Rep |
| Abstract |
Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by in vitro fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the tyrosinase gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3-47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats. |
| 巻・号 | 9(1) |
| ページ | 11571 |
| 公開日 | 2019-8-9 |
| DOI | 10.1038/s41598-019-47964-1 |
| PII | 10.1038/s41598-019-47964-1 |
| PMID | 31399630 |
| PMC | PMC6689013 |
| MeSH | Animals CRISPR-Cas Systems Electroporation / methods Electroporation / veterinary Female Fertilization in Vitro / methods Fertilization in Vitro / veterinary Gene Editing / methods Gene Editing / veterinary Gene Knock-In Techniques* / methods Gene Knock-In Techniques* / veterinary Gene Knockout Techniques* / methods Gene Knockout Techniques* / veterinary Male Rats / embryology* Rats / genetics Rats / physiology Rats, Inbred F344 / embryology Rats, Inbred F344 / genetics Rats, Inbred F344 / physiology Rats, Long-Evans / embryology Rats, Long-Evans / genetics Rats, Long-Evans / physiology Rats, Sprague-Dawley / embryology Rats, Sprague-Dawley / genetics Rats, Sprague-Dawley / physiology Rats, Wistar / embryology Rats, Wistar / genetics Rats, Wistar / physiology Superovulation |
| IF | 4.011 |
| 引用数 | 4 |
| リソース情報 | |
| ラット | NBRP-Ratの基盤整備プログラムの研究に関する論文 |