2021 Fiscal Year Final Research Report
Development of bioorthogonal RNA-RNA-binding protein pairs via molecular co-evolution and application for translational control
Project/Area Number |
19K15701
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Research Category |
Grant-in-Aid for Early-Career Scientists
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Allocation Type | Multi-year Fund |
Review Section |
Basic Section 37010:Bio-related chemistry
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Research Institution | Okinawa Institute of Science and Technology Graduate University |
Principal Investigator |
FUKUNAGA KEISUKE 沖縄科学技術大学院大学, 核酸化学・工学ユニット, ポストドクトラルスカラー (80639279)
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Project Period (FY) |
2019-04-01 – 2022-03-31
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Keywords | PD-SELEX / RNAアプタマー / RNA結合タンパク質 / 進化分子工学 / 共進化 |
Outline of Final Research Achievements |
A novel library-vs-library in vitro selection method (PD-SELEX: Phage Display coupled with Systematic Evolution of Ligands by EXponential enrichment) has been developed. Starting with pools of 1.1 x 10^12 unique RNA sequences and 4.0 x 10^8 unique phage-displayed L7Ae-Scaffold (LS) proteins, six rounds of selection were carried out. The selected RNAs and LS proteins were analyzed by high-throughput sequencing. Further deconvolution of the enriched RNA and LS protein sequences revealed two orthogonal RNA-RBP pairs that exhibit 7 pM KD value and >4000-fold selectivity.
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Free Research Field |
生体分子工学
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Academic Significance and Societal Importance of the Research Achievements |
核酸-タンパク質(ペプチド)間相互作用を解析、または結合選択性を改変するための基盤技術としてPD-SELEX法が活用できる。また、セレクションで得られた直交性RNA-RNA結合タンパク質ペア(CS1 RNA-LS4 protein & CS2 RNA-LS12 protein)は、任意のRNAとタンパク質を繋ぐアダプター分子としての利用が見込まれる。
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