202003Nakagawa

研究発表を行った学会；日本ゲノム編集学会　第4回大会
2019年 6月 3日〜 5日（東京）
タイトル； Highly versatile generation of electroporation-mediated genome-edited mice via the CREATRE method.
発表者；中川　佳子氏
（熊本大学　生命資源研究・支援センター　資源開発分野）
要旨；
Recently, we reported on the efficient and quick production of electroporation-mediated genome-edited mice using the CREATRE (CARD-based Reproductive Engineering-Assisted Technology for RNP Electroporation) method. The CREATRE method offers superior work efficiency and flexible scheduling by combining genome editing technology with reproductive engineering techniques such as ultra-superovulation, in vitro fertilization (IVF), and the vitrification/warming of zygotes. In this study, to preferentially obtain non-mosaic mutants, we investigated the suited condition of electroporation to generate mutant mice presenting single-peak mutation by direct sequencing analysis, indicating homozygously mutated mice. By altering the voltage of electroporation and the culture time after the warming of zygotes, we identified the optimal protocol for the creation of mutant mice carrying single-peak mutations. Furthermore, our protocol yielded an improved birth rate without decreasing the mutation rate in comparison to the previous CREATRE protocol, thereby further improving the efficiency of CREATRE-mediated generation of genome-edited mice.