Abstract：
Amplifying DNA conjugated affinity ligands can improve the sensitivity and multiplicity of cell imaging and play a crucial role in comprehensively deciphering cellular heterogeneity and dynamic changes during development and disease. However, the development of one-step, controllable, and quantitative DNA amplification methods for multiplexed imaging of live-cell membrane proteins is challenging. Here, we introduce the template adhesion reaction (TAR) method for assembling amplifiable DNA sequences with different affinity ligands, such as aptamers or antibodies, for amplified and multiplexed imaging of live-cell membrane proteins with high quantitative fidelity. The precisely controllable TAR enables proportional amplification of membrane protein targets with variable abundances by modulating the concentration ratios of hairpin templates and primers, thus allowing sensitive visualization of multiple membrane proteins with enhanced signal-to-noise ratios (SNRs) without disturbing their original ratios. Using TAR, we achieved signal-enhanced imaging of six proteins on the same live-cell within 1-2 h. TAR represents an innovative and programmable molecular toolkit for multiplexed profiling of membrane proteins in live-cells.
摘要：
扩增DNA亲和配体可以提高细胞成像的敏感性和多重性，在全面破译发育和疾病过程中的细胞异质性和动态变化中起着至关重要的作用。然而,一步到位的发展,可控,和用于活细胞膜蛋白多重成像的定量DNA扩增方法具有挑战性。这里,我们介绍了模板粘附反应（TAR）方法，用于组装具有不同亲和配体的可扩增DNA序列，如适体或抗体，用于具有高定量保真度的活细胞膜蛋白的扩增和多重成像。精确可控的TAR可以通过调节发夹模板和引物的浓度比来实现可变丰度的膜蛋白靶标的成比例扩增。因此，可以对具有增强的信噪比（SNR）的多种膜蛋白进行灵敏的可视化，而不会干扰其原始比率。使用TAR,我们在1-2小时内实现了6种蛋白质在同一活细胞上的信号增强成像。TAR代表了一种创新和可编程的分子工具包，用于活细胞中膜蛋白的多重分析。
