Production of animal population having identical modified gene in a short period by genome editing and nuclear transplantation techniques in Xenopus leaves.

Research Project

Project/Area Number	15K14562
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Morphology/Structure
Research Institution	Kumamoto University
Principal Investigator	Takamune Kazufumi 熊本大学, 大学院先端科学研究部(理), 教授 (20206882)
Co-Investigator(Kenkyū-buntansha)	北野健熊本大学, 大学院先端科学研究部(理), 准教授 (40336219)
Project Period (FY)	2015-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000) Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywords	ゲノム編集 / 核移植 / クローン / 凍結保存 / Xenopus leavis / 両生類
Outline of Final Research Achievements	Genome editing techniques such as CRISPR/Cas9 system enable us to analyze the gene function. However, production of F1 (sometimes F2) generation is necessary for confirming the gene function by repetitive analyses because the state of mutations is different among the genome-edited individuals even if they are produced by the same procedure. In animals that need long periods for becoming sexually mature, we have to take long time to get population having the same mutation in the target gene. To overcome this disadvantage, I produced clonal population of genome-edited Xenopus laevis by transplantation of nucleus from embryo genome-edited by CRISPR/Cas9 system into enucleated egg. In addition, I succeeded in transplantation of nucleus from embryo which had been cryopreserved by vitrification. This technique allows us not only to ensure a sufficient time for selection of embryo having the desired mutation in the target gene but also to keep the useful animal in the narrow space.