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由混杂的RNA Aptamer和红色或蓝色化染料组成的模块:选择,特征和生物成像

Xiaohong Tan¹, Tudor P Constantin¹, Kelly L Sloane¹

¹Departments of Chemistry and Biological Sciences, Molecular Biosensor and Imaging Center, and Center for Nucleic Acids Science and Technology, Carnegie Mellon University , 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-3890, United States.

Journal of the American Chemical Society

|June 24, 2017

概括

这项研究介绍了一种多功能RNA吸收体,它结合了两种不同的色素,使双色光成像成为可能. 这种双功能性受体还可以准细胞表面受体,如表皮生长因子受体 (EGFR),用于现场成像和跟踪.

科学领域:

分子生物学
生物技术
化学生物学

背景情况:

核糖核酶是选择特定结合目标的核酸分子.
在生物成像中广泛使用的色剂.
开发具有可调节性质的新型成像探测器对于细胞生物学至关重要.

研究的目的:

开发一种具有双色光能力的新型RNA光体.
创建一个针对细胞表面受体标记的双功能合体.
为了证明aptamer在区分细胞表面和内化受体中的实用性.

主要方法:

通过指数式丰富 (SELEX) 来选择色素染料的系统演化.
聚胺染料结合动力学 (K_D) 和光物理性质 (量子产量,光稳定性) 的表征.
与表皮生长因子受体 (EGFR) 特定的染料结合剂的融合.

主要成果:

鉴定了一种单一的RNA胺基,它与二甲红色 (DIR) 和氧蓝色 (OTB) 结合,具有亚微分子亲和力.
由此产生的胺染料复合物 (模块) 具有可调节的光发射和高光稳定性.
在哺乳动物细胞上成功标记细胞表面EGFR.
通过改变结合的染料实现了光色 (蓝色/红色) 的现场切换.
该系统允许根据染料添加时间区分细胞表面和内化EGFR.

结论:

杂乱的RNA体为双色光成像提供了一个多功能平台.
这种双功能阿普坦能够对细胞表面受体进行有针对性的标记和动态跟踪.
这项技术为区分活细胞中的受体定位和动态提供了一种新的方法.

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