向m6A读者蛋白研究单个RNA的表谱调节
Journal of the American Chemical Society
|September 6, 2018
概括
研究人员开发了可编程的RNA结合蛋白来研究活细胞中的N6-甲基氨酸 (m6A) 修饰. 这种工具可以精确控制特定转录的RNA稳定性和翻译.
科学领域:
- 分子生物学
- 表观遗传学
- 核糖核酸生物学
背景情况:
- 转录后的基因调节对于哺乳动物的蛋白质生产至关重要.
- 化学修饰RNA,如N6-甲基氨酸 (m6A),是关键的调节机制.
- 在单个转录层面的m6A调节器 (写入器,擦除器,读取器) 的相互作用仍然不清楚.
研究的目的:
- 开发一种工具,用于研究特定的m6A阅读器对活细胞单个RNA转录的调节作用.
- 阐明m6A调节器在控制RNA命运中的竞争性相互作用.
主要方法:
- 通过将触媒不活性的PspCas13b与m6A读取蛋白 (YTHDF1,YTHDF2) 融合,开发可编程的dPspCas13b-m6A读取蛋白.
- 使用导向RNA (gRNA) 来针对性地将读者蛋白传递给特定感兴趣的RNA.
- 在记者结构和内源mRNA标上证明读者蛋白的功能.
主要成果:
- 融合读者蛋白 (YTHDF2,YTHDF1) 保留了它们已知的功能:YTHDF2诱导RNA降解,YTHDF1增强翻译.
- 该系统成功准了内源mRNA转录.
- 将YTHDF2与内源mRNA结合导致其衰变.
结论:
- 基于dPspCas13b的工具可以对活细胞中的RNA调节进行特异性研究.
- 这些工具增强了对单个转录水平的RNA调节的理解.
- 开发的工具在合成生物学中具有潜在的应用,用于通过RNA操纵控制基因表达.
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