Introduction of chromosome structural changes into mouse spermatogonia cells for the analysis of their transmission to next generation

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K12179
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 63020:Radiation influence-related
• Research Institution: Radiation Effects Research Foundation
• Principal Investigator: NODA Asao 公益財団法人放射線影響研究所, 分子生物科学部, 部長 (40294227)
• Co-Investigator(Kenkyū-buntansha): 濱崎 幹也 公益財団法人放射線影響研究所, 分子生物科学部, 研究員 (80443597)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 放射線 / 突然変異 / 生殖細胞変異 / 遺伝子編集 / 染色体構造変異 / CRISPR/Cas9 / 精原細胞

Outline of Final Research Achievements

We tried to create chromosomal structural mutations in the cultured cells. This is the creation of artificial chromosomal structural mutants by assuming characteristic mutations caused by radiation exposure. We investigate the possibility of inheritance of germ cells bearing these mutations to the next generation. This is an application of the CRISPR/Cas9-based gene-editing system to generate two double-strand breaks at specific sites on the mouse chromosome and create a deletion mutation between them by non-homologous end joining (NHEJ). As a result, we have successfully created extensive artificial deletions ranging from 100 bp to 1 Mbp. We then attempted to create artificial chromosomal translocations.

Free Research Field

放射線生物学、放射線遺伝学

Academic Significance and Societal Importance of the Research Achievements

放射線被ばくの遺伝的影響は生殖系列細胞に生じる突然変異に起因する。どのようなタイプの突然変異がどれくらいの効率で次世代に伝わるか、これまで明らかになっていない。体細胞とは異なり、生殖系列細胞には減数分裂チェックポイントや受精および初期発生という関門があり、これをくぐり抜けた細胞のみが次世代へと伝わる。生体内のそれぞれの過程で、染色体構造変異を持つ細胞がどのように取り扱われるのか、本研究はその検証システム作成を目指すものであり、生殖細胞被ばくのリスク推定の礎となると期待する。