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Title: Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization
Authors: Honda, Arata
Tachibana, Ryoma
Hamada, Kazuya
Morita, Kohtaro
Mizuno, Naoaki
Morita, Kento
Asano, Masahide
Author's alias: 本多, 新
橘, 亮麿
濱田, 和弥
守田, 昂太郎
水野, 直彬
森田, 健斗
浅野, 雅秀
Keywords: Development
Genetic engineering
Rat
Issue Date: 9-Aug-2019
Publisher: Springer Nature
Journal title: Scientific Reports
Volume: 9
Thesis number: 11571
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.
Description: 「誰でも」「簡単に」「効率よく」モデルラットを作出できる技術を開発. 京都大学プレスリリース. 2019-08-22.
Rights: © The Author(s) 2019. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
URI: http://hdl.handle.net/2433/243553
DOI(Published Version): 10.1038/s41598-019-47964-1
PubMed ID: 31399630
Related Link: http://www.kyoto-u.ac.jp/ja/research/research_results/2019/190809_1.html
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