DNA nanotechnology is an emerging field that promises fascinating opportunities for the manipulation and imaging of proteins on a cell surface. The key to progress is the ability to create a nucleic acid–protein junction in the context of living cells. Here we report a covalent labelling reaction that installs a biostable peptide nucleic acid (PNA) tag. The reaction proceeds within minutes and is specific for proteins carrying a 2 kDa coiled-coil peptide tag. Once installed, the PNA label serves as a generic landing platform that enables the recruitment of fluorescent dyes via nucleic acid hybridization. We demonstrate the versatility of this approach by recruiting different fluorophores, assembling multiple fluorophores for increased brightness and achieving reversible labelling by way of toehold-mediated strand displacement. Additionally, we show that labelling can be carried out using two different coiled-coil systems, with epidermal growth factor receptor and endothelin receptor type B, on both HEK293 and CHO cells. Finally, we apply the method to monitor internalization of epidermal growth factor receptor on CHO cells.

DNA 纳米技术是一个新兴领域，它为细胞表面蛋白质的操作和成像提供了迷人的机会。取得进展的关键是在活细胞环境中创建核酸-蛋白质连接的能力。在这里，我们报告了安装生物稳定肽核酸 (PNA) 标签的共价标记反应。该反应在几分钟内进行，并且对带有 2 kDa 卷曲螺旋肽标签的蛋白质具有特异性。安装后，PNA 标签可用作通用着陆平台，可通过核酸杂交募集荧光染料。我们通过招募不同的荧光团、组装多个荧光团以增加亮度并通过立足点介导的链置换实现可逆标记来证明这种方法的多功能性。此外，我们表明，可以在 HEK293 和 CHO 细胞上使用两种不同的螺旋线圈系统进行标记，即表皮生长因子受体和 B 型内皮素受体。最后，我们应用该方法监测CHO细胞表皮生长因子受体的内化。