2022 Fiscal Year Final Research Report
Development of methodology for subtractive design of microbiota
Project/Area Number |
20K21275
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Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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Allocation Type | Multi-year Fund |
Review Section |
Medium-sized Section 38:Agricultural chemistry and related fields
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Research Institution | Osaka University |
Principal Investigator |
Honda Kohsuke 大阪大学, 生物工学国際交流センター, 教授 (90403162)
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Project Period (FY) |
2020-07-30 – 2023-03-31
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Keywords | 菌叢解析 / ペプチド核酸 / 細胞膜透過ペプチド |
Outline of Final Research Achievements |
In this study, we aimed at the development of a novel molecular tool which can induce cell death only to specific genera and species of bacteria among various bacteria composing of a microbial consortium (microbiota). We first tried to inactivate an essential gene of target bacteria by delivering a Cas nuclease into the cell using cell penetration peptides (CPP). However, we could not observe the penetration of the CPP-fused Cas nuclease into the cells probably due to the large size of the fusion protein. Accordingly, we employed peptide nucleic acid (PNA), which is a much smaller molecule than Cas nuclease, and introduced them in bacterial cells using CPP. Consequently, we confirmed that a 2-8 μM of CPP-fused PNA induced specific cell death to Escherichia coli and Pseudomonas putida in artificial microbiota.
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Free Research Field |
応用微生物学
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Academic Significance and Societal Importance of the Research Achievements |
ヒト共生微生物群集研究の発展などを契機に、近年、微生物群集(菌叢)の機能に注目が集まっている。当該分野では、純粋培養した微生物種を組み合わせて作成された人工菌叢を用いる再構成的研究によって、一定の成果が得られつつあるが、減算的アプローチによる菌叢解析技術、すなわち既存の既存の菌叢から特定の微生物種だけを取り除くための有効な方法論が存在しなかった。本研究で開発されたCPP融合ペプチド核酸による選択的細胞死誘導技術は、こうした現状を打破する基盤技術のひとつと位置づけられ、菌叢中の微生物間相互作用の直接的観察を可能にするなど、当該分野に大きなインパクトをもたらすものとなりうる。
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