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快速交换的螺旋环罗达胺探针用于基于aptamer的超分辨率RNA成像

Daniel Englert¹, Eva-Maria Burger¹, Franziska Grün¹

¹Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany.

Nature communications

|June 30, 2023

概括

研究人员开发了RhoBAST:SpyRho,这是一种用于高分辨率活细胞RNA成像的新型光照明吸收器 (FLAP) 系统. 这个系统克服了以前的局限性,使RNA和其他细胞组件的清晰可视化.

科学领域:

分子生物学分子生物学
生物物理学的生物物理.
显微镜的使用方法

背景情况:

活细胞RNA成像需要高空间和时间分辨率,这是细胞生物学中持续存在的挑战.
现有的光探测器往往受到细胞透性差,亮度低,信号与背景比不够的困扰.

研究的目的:

开发一种先进的光照明吸收器 (FLAP) 系统,用于优异的活细胞RNA可视化.
克服当前探针的局限性,并使先进的光显微镜技术成为可能.

主要方法:

开发了一种新型的探针SpyRho (Spirocyclic Rhodamine),该探针旨在与RhoBAST体结合高亲和力.
利用螺旋和化物形式之间的平衡转移来增强亮度和性.
在超分辨率显微镜技术中的应用,包括超分辨率单分子定位显微镜 (SMLM) 和刺激辐射耗尽 (STED) 显微镜.

主要成果:

罗巴斯特:SpyRho显示出高亮度,性,以及信号与背景的比率.
在活哺乳动物细胞中实现了第一个特别标记的RNA的超高分辨率STED图像.
成功可视化了内源性染色体位置和蛋白质,展示了系统的多功能性.

结论:

罗巴斯特:SpyRho代表了活细胞RNA成像的重大进步,超过了现有的FLAP系统.
该系统在SMLM和STED成像中的性能为研究RNA动态和定位开辟了新的途径.
它的多功能性扩展到成像其他细胞目标,突出其在细胞生物学研究中的广泛适用性.

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