宿主トランスクリプトから派生した非正規のcrRNAは,Cas9によって複合性RNAの検出を可能にします
Chunlei Jiao1, Sahil Sharma2, Gaurav Dugar2
1Helmholtz Institute for RNA-based Infection Research (HIRI)/Helmholtz-Centre for Infection Research (HZI), 97080 Würzburg, Germany.
Science (New York, N.Y.)
|April 28, 2021
まとめ
科学者は非正規のCRISPRRNAs (crRNAs) を作成することで,RNAを検出するCRISPR-Casシステムを設計しました. これにより,SARS-CoV-2の変種を含む複合RNA検出のためのLEOPARD診断プラットフォームが開発されました.
科学分野:
- 分子生物学
- CRISPR-Casシステム
- 診断技術
背景:
- CRISPR-Casシステムは,外来遺伝物質を識別するためにCRISPR RNAs (crRNAs) を使用しています.
- タイプIIのCRISPR-Casシステムは,crRNA処理とCas9酵素利用のためにトランス活性化crRNA (tracrRNA) に依存しています.
- トラクルRNAは典型的には特定のcrRNAとハイブリッド化します.
研究 の 目的:
- カンピロバクター・ジェジュニのCas9-RNA複合体の相互作用を調査する.
- 新しいcrRNA形成メカニズムとDNAターゲティングの可能性を発見する.
- 改造されたtracrRNAをベースにした多機能RNA検出プラットフォームを設計する.
主な方法:
- カンピロバクター・ジェジュニのCas9-RNA複合体の分析
- トラクルRNAが内生的な細胞RNAにハイブリッド化し,非正規のcrRNAを形成する発見.
- Cas9媒介のDNAターゲティングとRNAの存在を結びつけるための再プログラムされたtracrRNAsのエンジニアリング.
- LEOPARDプラットフォームの開発 (並列RNA検出のためのエンジニアリングされたtracrRNAとオンターゲットDNAの活用)
主要な成果:
- Cas9 DNA ターゲティングを誘導する能力を持つ,細胞RNAへの tracrRNA ハイブリダイゼーションによって生成された非正規の crRNA を特定した.
- エンジニアリングされたtracrRNAは,任意の望ましいRNAの存在に基づいてDNAターゲティングを可能にしました.
- LEOPARDプラットフォームは,単一のテストで複数のウイルスRNAを同時に検出することができました.
- LEOPARDは,患者サンプルで単核酸精度でSARS-CoV-2とD614Gの変種を区別しました.
結論:
- 非正規のcrRNA形成の発見は,CRISPR-Casシステムの汎用性の理解を広げています.
- 設計されたtracrRNAはRNAによるDNAターゲティングのための新しいメカニズムを提供します.
- LEOPARDプラットフォームは,ウイルス診断を含む,敏感で特定のRNA検出のための強力なマルチプレクサブルツールを提供します.
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