TRIBE:劫持RNA编辑酶以识别RNA结合蛋白的细胞特异性目标
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
概括
研究人员开发了TRIBE,一种用于在体内识别RNA结合蛋白 (RBP) 的新方法. 这种技术甚至可以在小细胞群中精确识别目标,克服现有方法的局限性.
科学领域:
- 分子生物学
- 基因组学
- 生物化学
背景情况:
- 在调节RNA转录中,RNA结合蛋白 (RBPs) 起着至关重要的作用.
- 现有方法在体内识别RBP标是有限的,特别是对于小细胞数量.
研究的目的:
- 开发一种改进的技术,以在体内识别RNA结合蛋白的点.
- 在小,特定的细胞群中实现RBP目标识别.
主要方法:
- 开发了TRIBE (通过编辑识别的RNA结合蛋白的目标).
- TRIBE将一个RBP与DrosophilaRNA编辑酶ADAR的催化域结合起来.
- 通过标记新编辑事件的RNA测序来识别RBP目标.
主要成果:
- 使用TRIBE成功确定了三个RBP (Hrp48,dFMR1,NonA) 的目标.
- TRIBE的性能与CLIP等方法相美.
- 识别了150个特定的神经元中的RBP目标.
结论:
- TRIBE是一种有效的在体内识别RBP目标的技术.
- TRIBE克服了以前方法的局限性,可以在没有抗体的情况下对小细胞进行分析.
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