可编程,结构切换的RhoBAST用于活细胞中混合介导的mRNA成像
Bastian Bühler1, Janin Schokolowski1, Andres Jäschke1
1Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, 69120 Heidelberg, Germany.
ACS chemical biology
|August 2, 2023
概括
研究人员开发了可编程的RhoBAST吸收体,以可视化活细胞中未标记的信使RNA (mRNA). 这些探测器只在与特定的点RNA结合时才会发光,从而实现精确的RNA成像.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 细胞生物学 细胞生物学
背景情况:
- 在活细胞中可视化内源RNA对于理解它们的功能至关重要.
- 光发光光体 (FLAP) 为RNA成像提供了一种有前途的方法.
- 罗巴斯特是一种高度光稳定和明亮的FLAP,用于信使RNA (mRNA) 成像.
研究的目的:
- 设计可编程的RhoBAST序列,用于特定的,在现场检测未标记的目标RNA.
- 创建一个模块化系统,其中目标结合会诱导RhoBAST对光激活的形状变化.
- 为了证明可编程RhoBAST对活细菌mRNA可视化的实用性.
主要方法:
- 设计RhoBAST序列,并配一个包含模块化传感器的杂交臂.
- 结构切换和光照明的特征 in vitro.
- 可编程RhoBAST的应用用于细菌中未标记的mRNA的实时成像.
主要成果:
- 可编程的RhoBAST序列成功地被设计成在转基因中结合特定的点RNA.
- 一个模块化传感器序列控制RhoBAST结构,使得目标依赖的光.
- 该系统在与TMR-DN.相互作用后与向mRNA结合时显示了特定的光照明.
- 在活细菌细胞中成功可视化未被标记的mRNA.
结论:
- 可编程的RhoBAST体为成像内源性,未标记的RNA提供了一种新和特定的方法.
- 这种模块化系统提供了针对不同RNA分子的多功能性.
- 开发的探针适用于可视化活细菌系统中的mRNA动态.
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