Development of polycistronic gene expression system for yeast synthetic biology

• Project/Area Number: 18K14374
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 38020:Applied microbiology-related
• Research Institution: Kobe University
• Principal Investigator: Masahiro Tominaga 神戸大学, 先端バイオ工学研究センター, 特命助教 (50761409)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
• Keywords: 遺伝子発現システム / 酵母 / 翻訳 / 遺伝子発現 / 合成生物学 / リボソーム / 進化工学 / IRES

Outline of Final Research Achievements

Gene expression tools are fundamental in any biotechnological project. In this project, we established a gene expression system using an internal ribosomal entry site (IRES) to realize efficient polycistronic gene expression in eukaryotic yeast. We developed a screening system for the evaluation of IRES-dependent translation efficiency in yeast. By using this screening system, we could find several requirements for the efficient IRES-dependent translation in yeast. Leveraging the findings, further engineering of the IRES-dependent translation can be performed in yeast.

Academic Significance and Societal Importance of the Research Achievements

遺伝子発現は生物機能の根幹であり，その制御系の充実は生物機能の工学自由度に直結する。その点で，1つのRNAから1つの遺伝子しか発現できない真核生物では，原核生物に比べ遺伝子発現系構築の自由度が低い。本研究の目的は，真核生物の酵母に，1つのRNAから複数遺伝子を発現できる「オペロン型の遺伝子発現系」を構築することであり，人工遺伝子発現ネットワークの構築や有価物質生産のための代謝経路構築などを加速すると期待される。