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2023 Fiscal Year Final Research Report

Developing new model microorganisms with diverse nutritional strategies by enhancing growth efficiency through genetic manipulation

Research Project

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Project/Area Number 21K19240
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionFukui University of Technology

Principal Investigator

Kashiyama Yuichiro  福井工業大学, 環境学部, 教授 (00611782)

Co-Investigator(Kenkyū-buntansha) 藤原 崇之  国立遺伝学研究所, 遺伝形質研究系, 助教 (10595151)
中澤 昌美  大阪公立大学, 生命環境科学研究科, 講師 (90343417)
Project Period (FY) 2021-07-09 – 2024-03-31
KeywordsCRISPR/Cas9ゲノム編集 / 非モデル生物 / 難培養性 / ラパザ / 盗葉緑体現象
Outline of Final Research Achievements

In this study, we aimed to establish and apply a transformation system for non-model organisms, with the ultimate goal of achieving efficient cultivation by introducing cell membrane transporters to utilize dissolved organic matter such as sugars and amino acids absorbed from the culture medium. During this period, we developed an experimental system aimed at CRISPR/Cas9 gene knock-in, particularly for the phagotrophic mixotroph (thallophyte) Rapaza viridis (Rapaza), and achieved high-efficiency (up to 60%) genome editing, successfully conducting knockout experiments and experiments to introduce epitope tags. In addition, we identified the transporter protein of R. viridis that is expressed in the cell membrane, and proceeded with verification experiments for the introduction of protein genes fused to this.

Free Research Field

細胞生物学

Academic Significance and Societal Importance of the Research Achievements

人類が有する真核細胞の知識の大半は,自然界の広範な多様性の中のごく一部の「培養しやすい」モデル生物の研究に基づいている。よって多様な真核生物(細胞)の大半が実験室での培養が困難である事実を考えれば,我々の知識はむしろ非常に特殊な生物からの偏った情報に依存している可能性さえある。我々は,近年確立されたゲノム編集技術はこれら「人類の知らない細胞」の研究に革命をもたらすポテンシャルを有していると考えた。本研究は,ラパザという,これまで実験細胞として見向きもされず,その「盗葉緑体現象」という「常識」からすれば奇妙奇天烈な細胞生理を示す生物のゲノム編集実験系を確立し,新しい真核細胞研究をスタートさせた。

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Published: 2025-01-30  

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