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高速AFMによるゲノム編集ツールCRISPR-Casの分子作動機構の網羅的解析
https://doi.org/10.24517/00059078
https://doi.org/10.24517/00059078dbbfc61a-866e-4209-b78a-cbc5a1270237
名前 / ファイル | ライセンス | アクション |
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FR-PR-SHIBATA-M-kaken 2021-12p.pdf (993.3 kB)
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Item type | 報告書 / Research Paper(1) | |||||
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公開日 | 2022-03-03 | |||||
タイトル | ||||||
タイトル | 高速AFMによるゲノム編集ツールCRISPR-Casの分子作動機構の網羅的解析 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | Comprehensive studies of working mechanism of Cas proteins using HS-AFM | |||||
言語 | ||||||
言語 | jpn | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_18ws | |||||
資源タイプ | research report | |||||
ID登録 | ||||||
ID登録 | 10.24517/00059078 | |||||
ID登録タイプ | JaLC | |||||
著者別表示 |
Shibata, Mikihiro
× Shibata, Mikihiro |
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提供者所属 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 金沢大学ナノ生命科学研究所 | |||||
書誌情報 |
令和2(2020)年度 科学研究費補助金 基盤研究(B) 研究成果報告書 en : 2020 Fiscal Year Final Research Report 巻 2018-04-01 – 2021-03-31, p. 12p., 発行日 2021-05-14 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | 本研究は、ゲノム編集ツールCasタンパク質群に高速AFMを適用し、様々なCasタンパク質がDNAに結合し切断する瞬間を実時空間で可視化することで、DNA切断分子作動機構を統一的に理解することを目的とした。研究期間において、SaCas9、AsCas12a、LbCas12aに対して高速AFM観察を行い、核酸非結合状態ではフレキシブルな構造をとるが、RNA結合状態では安定で固い構造をとり、RNA-DNA結合状態では、標的DNA配列に強く結合したままであることを明らかにした。この分子動態は、全てのCasタンパク質において観察され、エンドヌクレアーゼにおける共通の分子作動機構といえる。 | |||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | The purpose of this study was to understand the mechanism of DNA cleavage reaction of Cas proteins, which are used as genome editing tools. Using high-speed atomic force microscopy (HS-AFM), we tried to visualize the dynamics of Cas proteins during the binding to and cleaving a target site of DNA in real-time with nano-meter resolution. During the three years research period, HS-AFM observations were performed on SaCas9, AsCas12a, and LbCas12a. HS-AFM videos of three Cas proteins showed that structures of Cas proteins without nucleic acids were flexible, while structures of RNA-binding Cas proteins were stable and rigid. Furthermore, when RNA-Cas complex bound to the target site of DNA, a complex remained tightly bound to the target site of DNA and did not slide on the DNA. Since this molecular dynamics was observed in all Cas proteins, we concluded that it could be a common molecular binding mechanism to the target site of DNA of Cas proteins. | |||||
内容記述 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 研究課題/領域番号:18H01836, 研究期間(年度):2018-04-01 – 2021-03-31 | |||||
内容記述 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 出典:「高速AFMによるゲノム編集ツールCRISPR-Casの分子作動機構の網羅的解析」研究成果報告書 課題番号18H01836 (KAKEN:科学研究費助成事業データベース(国立情報学研究所)) (https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-18H01836/18H01836seika/)を加工して作成 |
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著者版フラグ | ||||||
出版タイプ | AM | |||||
出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/ja/search/?kw=80631027 | |||||
関連名称 | https://kaken.nii.ac.jp/ja/search/?kw=80631027 | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-18H01836/ | |||||
関連名称 | https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-18H01836/ | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-18H01836/18H01836seika/ | |||||
関連名称 | https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-18H01836/18H01836seika/ |