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Directed evolution of orthogonal RNA–RBP pairs through library-vs-library in vitro selection
https://oist.repo.nii.ac.jp/records/2601
https://oist.repo.nii.ac.jp/records/260119a1673e-1f24-4da1-9148-c81ca2189b2f
名前 / ファイル | ライセンス | アクション |
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Fukunaga-2022-Directed evolution of orthogonal (3.4 MB)
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Creative Commons Attribution 4.0 International(https://creativecommons.org/licenses/by/4.0/)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2022-03-23 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | Directed evolution of orthogonal RNA–RBP pairs through library-vs-library in vitro selection | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者(英) |
Fukunaga, Keisuke
× Fukunaga, Keisuke× Yokobayashi, Yohei |
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書誌情報 |
en : Nucleic Acids Research 巻 50, 号 2, p. 601-616, 発行日 2021-07-05 |
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抄録 | ||||||
内容記述タイプ | Other | |||||
内容記述 | RNA-binding proteins (RBPs) and their RNA ligands play many critical roles in gene regulation and RNA processing in cells. They are also useful for various applications in cell biology and synthetic biology. However, re-engineering novel and orthogonal RNA–RBP pairs from natural components remains challenging while such synthetic RNA–RBP pairs could significantly expand the RNA–RBP toolbox for various applications. Here, we report a novel library-vs-library in vitro selection strategy based on Phage Display coupled with Systematic Evolution of Ligands by EXponential enrichment (PD-SELEX). Starting with pools of 1.1 × 10¹² unique RNA sequences and 4.0 × 10⁸ unique phage-displayed L7Ae-scaffold (LS) proteins, we selected RNA–RBP complexes through a two-step affinity purification process. After six rounds of library-vs-library selection, the selected RNAs and LS proteins were analyzed by next-generation sequencing (NGS). Further deconvolution of the enriched RNA and LS protein sequences revealed two synthetic and orthogonal RNA–RBP pairs that exhibit picomolar affinity and >4000-fold selectivity. | |||||
出版者 | ||||||
出版者 | Oxford University Press on behalf of Nucleic Acids Research | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 1362-4962 | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 0305-1048 | |||||
PubMed番号 | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | PMID | |||||
関連識別子 | info:pmid/34219162 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | info:doi/10.1093/nar/gkab527 | |||||
権利 | ||||||
権利情報 | © 2021 The Author(s). | |||||
関連サイト | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://academic.oup.com/nar/article/50/2/601/6313240 | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 |