Establishment of stable somatic haploid cells through engineering of centrosome duplication "checkpoint"

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K19244
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 44:Biology at cellular to organismal levels, and related fields
• Research Institution: Hokkaido University
• Principal Investigator: Uehara Ryota 北海道大学, 先端生命科学研究院, 准教授 (20580020)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Keywords: 倍数性 / 中心体 / 一倍体

Outline of Final Research Achievements

In this study, we attempted to resolve the genomic instability of animal haploid cells caused by chronic centrosome loss using two approaches: i) designing a new molecular circuit to resolve centrosome replication defects and ii) establishing a genetic background that allows stable mitotic control in the absence of a centrosome. Regarding i), we examined and optimized the structure of the artificial circuit component genes and produced synthetic genes that minimize interference with the endogenous centrosome regulatory system. We will evaluate their functions in the future. For ii), we found haploidy-linked insufficiency in gene products for acentrosomal mitotic control. By the replenishment of these factors, we succeeded in establishing a stable haploid cell line that could maintain their haploid state for more than one month in continuous culture.

Free Research Field

細胞生物学

Academic Significance and Societal Importance of the Research Achievements

全遺伝子を１コピー保有する一倍体細胞は、遺伝子機能改変が容易で、遺伝学や細胞工学ツールとして高い利便性をもつ。しかし、動物一倍体は著しいゲノム不安定性で短期間に二倍体化することが汎用性の妨げている。本研究では、二つの独立したアプローチのうちの１つにより高い安定性をもつ一倍体ヒト細胞の樹立に成功した。この過程で、一倍体不安定性の原因となる具体的な遺伝子群を特定した点に高い学術的意義がある。また、本研究で確立した一倍体安定化法は、多くの動物種の一倍体制御に適用可能と考えられ、安定一倍体技術の発達を通して、生命工学および農水産資源開発の分野に波及性が期待できる点に社会的意義がある。