TRIBE:RNA結合タンパク質の細胞特異的標的を特定するためにRNA編集酵素をハイジャックする
Aoife C McMahon1, Reazur Rahman1, Hua Jin1
1Department of Biology, Howard Hughes Medical Institute and National Center for Behavioral Genomics, Brandeis University, Waltham, MA 02453, USA.
Cell
|April 5, 2016
まとめ
研究者は,RNA結合タンパク質 (RBP) の標的を in vivo で特定するための新しい方法である TRIBE を開発しました. この技術は,小さな細胞集団でも正確な標的の識別を可能にし,既存の方法の限界を克服します.
科学分野:
- 分子生物学
- ゲノミクス
- 生物化学
背景:
- RNA結合タンパク質 (RBPs) は,RNAの転写を調節する上で重要な役割を果たします.
- RBPの標的を in vivoで特定するための既存の方法は,特に小さな細胞数には限られています.
研究 の 目的:
- RNA結合タンパク質の in vivo 標的を特定するための改良された技術を開発する.
- 小さな特定の細胞集団での RBP 標的の識別を可能にします.
主な方法:
- 開発されたTRIBE (編集によって特定されたRNA結合タンパク質の標的).
- TRIBEはRBPをドロソフィラRNA編集酵素ADARの触媒ドメインと結合する.
- RBPの標的は,新しい編集イベントでマークされたRNAのシーケンスによって識別されます.
主要な成果:
- TRIBEを使用した3つのRBP (Hrp48,dFMR1,NonA) のターゲットが成功しました.
- TRIBEはCLIPのような方法と同等またはそれより優れた性能を示した.
- 特定されたRBPの標的は 150の特定のフライニューロンから
結論:
- TRIBEは,RBPの標的をインビボで特定するための効果的な技術です.
- TRIBEは以前の方法の限界を克服し,抗体なしで小さな細胞数での分析を可能にします.
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