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通过精确可控的DNA编码放大反应对活细胞膜蛋白质进行多重化和定量成像

Nannan Diao^1,2, Jianing Hou^1,2, Xinyu Peng^1,3

¹Institute of Molecular Medicine and Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine, State Key Laboratory of Systems Medicine for Cancer, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200127, China.

Angewandte Chemie (International ed. in English)

|July 9, 2024

概括

我们开发了一种新的DNA放大方法,称为模板粘附反应 (TAR),以改善活细胞成像. TAR允许同时对多个膜蛋白进行敏感,定量和多重可视化.

科学领域:

分子生物学分子生物学
细胞生物学细胞生物学
生物技术是生物技术.

背景情况:

用DNA增强的亲和关系联体增强了细胞成像灵敏度和多重性.
解读细胞异质性和发育和疾病的动态需要先进的成像.
对多重活细胞成像的可控,定量,单步DNA放大仍然是一个挑战.

研究的目的:

介绍模板粘附反应 (TAR) 方法,用于组装具有亲缘关系联结体的可放大DNA序列.
为了使活细胞膜蛋白质的放大和多重成像能够具有高定量准确性.
开发一种可控制和可编程的分子工具包,用于多重膜蛋白质分析.

主要方法:

使用 TAR 方法将可放大 DNA 序列与亲和性联结体 (吸收体或抗体) 组合起来.
通过调节发针模板和原料度以获得比例放大,精确控制TAR.
在不改变它们的原始比率的情况下,实现多重膜蛋白的信号增强成像.

主要成果:

TAR可使膜蛋白点的比例放大,其丰度可变.
通过增强的信号噪声比率 (SNRs) 实现了多重膜蛋白的敏感可视化.
通过信号增强成像,在1-2小时内在同一个活细胞上对六种蛋白质进行成像.

结论:

TAR是一种创新的,可编程的分子工具包,用于活细胞中的膜蛋白质的多重分析.
该方法允许对细胞异质性和动态进行敏感和定量成像.
在多重活细胞成像中,TAR克服了单步可控DNA放大方面的挑战.

关键词:

放大和复杂成像成像的扩大和复杂成像.活细胞膜蛋白质的活细胞膜蛋白质.的比例放大放大.定量成像技术定量成像技术模板粘附反应反应模板

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