Identification and analysis of the susceptibility gene for streptozotocin-induced diabetes in mice using the genome editing technology

Research Project

Project/Area Number	17K14975
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Laboratory animal science
Research Institution	Nagoya University
Principal Investigator	Miyasaka Yuki 名古屋大学, 医学系研究科, 助教 (30778098)
Project Period (FY)	2017-04-01 – 2020-03-31
Project Status	Completed (Fiscal Year 2019)
Budget Amount *help	¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000) Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000) Fiscal Year 2017: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywords	マウス / ストレプトゾトシン / 糖尿病感受性遺伝子 / 膵島脆弱性 / ゲノム編集技術 / ゲノム編集 / CRISPR/Cas9システム / 遺伝学 / CRISPR/Cas9 / 体系的遺伝解析系
Outline of Final Research Achievements	A missense mutation (p.Ala132Ser) in N-methylpurine-DNA glycosylase (Mpg) is a candidate mutation for streptozotocin (STZ) susceptibility in A/J mice. Alternatively, a missense mutation (p.Gly200Val) in DNA repair protein RAD50 (Rad50) is a candidate mutation for STZ susceptibility in NSY/Hos (NSY) mice. To confirm the effects of these mutations on STZ susceptibility, we developed a knock-in mice strain in which each mutation was replaced with a normal allele via CRISPR/Cas9-mediated genome editing technology. We analyzed STZ susceptibility in the knock-in mice and revealed that a p.Ala132Ser mutation in Mpg was not associated with STZ susceptibility in A/J mice. Conversely, a p.Gly200Val mutation in Rad50 was confirmed as a mutation that regulates STZ susceptibility in NSY mice.
Academic Significance and Societal Importance of the Research Achievements	マウス系統間にはSTZ感受性に差異が存在する事が知られているが、その原因遺伝子を同定した例は無い。本研究では、効果の小さい変異ではあるもののSTZ感受性を規定する遺伝子変異を特定することに成功した。STZ感受性遺伝子は膵島の脆弱性に関与していることが推定される為、本研究で特定した変異は膵島脆弱性に起因する糖尿病発症機構の解明に繋がるヒントになると期待される。