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SiRA:一种用于活细胞超分辨率RNA成像的罗达胺结合性吸收剂
Regina Wirth1, Peng Gao2,3, G Ulrich Nienhaus2,3,4,5
1Institute of Pharmacy and Molecular Biotechnology (IPMB) , Heidelberg University , 69120 Heidelberg , Germany.
Journal of the American Chemical Society
|April 16, 2019
概括
研究人员开发了一种新型的RNA吸收酶 (SiRA),可以结合罗达胺 (SiR) 染料,从而使活细胞RNA成像具有明亮,光稳定的远红色光辐射. 该系统使用超分辨率显微镜对细菌和活细胞中的RNA进行了增强的可视化.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 生物物理学的生物物理.
背景情况:
- 遗传编码的亮光RNA吸收体对活细胞RNA可视化非常有价值.
- 远红色和近红外 (NIR) 发射的aptamer/ligand对是稀有的.
- 罗达胺 (SiRs) 是光稳定的,具有高细胞透性的NIR发射光体.
研究的目的:
- 开发一种用于远红/NIR光成像的新型照射RNA吸收体.
- 使用罗达胺染料创建一个多功能RNA标签工具.
- 展示该系统在活细胞RNA成像和超分辨率显微镜中的应用.
主要方法:
- 在实验室中从组合图书馆中选择RNA体.
- 生物信息分析,截断和突变研究以确定最小的体 (SiRA).
- 光物理特征和活细胞成像实验,包括STED显微镜.
主要成果:
- 一个50核酸最小的阿帕特马,SiRA,被确定为SiRs的纳米分子亲和力.
- SiRA具有很高的光稳定性,量子产量为0.98,并且灭绝系数为86,000 M-1cm-1.1.
- 使用SiRA成功可视化细菌中的RNA和活细胞中的mRNA,包括STED超分辨率成像.
结论:
- 到目前为止,SiRA-SiR系统是迄今为止报道的最明亮的远红色亮光光体.
- 这项工作是首次将SiR染料和分子内螺旋环化应用于基于aptamer的RNA成像.
- SiRA系统具有通过新型RNA标记技术推动生物研究的巨大潜力.
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