Department of Chemistry, University of Massachusetts, Amherst, MA, 01003, USA.
Institute for Applied Life Sciences, University of Massachusetts, Amherst, MA, 01003, USA.
Angew Chem Int Ed Engl. 2022 Feb 1;61(6):e202112033. doi: 10.1002/anie.202112033. Epub 2021 Dec 22.
The cell membrane is a dynamic and heterogeneous structure composed of distinct sub-compartments. Within these compartments, preferential interactions occur among various lipids and proteins. Currently, it is still challenging to image these short-lived membrane complexes, especially in living cells. In this work, we present a DNA-based probe, termed "DNA Zipper", which allows the membrane order and pattern of transient interactions to be imaged in living cells using standard fluorescence microscopes. By fine-tuning the length and binding affinity of DNA duplex, these probes can precisely extend the duration of membrane lipid interactions via dynamic DNA hybridization. The correlation between membrane order and the activation of T-cell receptor signaling has also been studied. These programmable DNA probes function after a brief cell incubation, which can be easily adapted to study lipid interactions and membrane order during different membrane signaling events.
细胞膜是一种由不同亚区室组成的动态、不均一的结构。在这些区室中，各种脂质和蛋白质之间会发生优先相互作用。目前，仍然难以对这些短暂存在的膜复合物进行成像，特别是在活细胞中。在这项工作中，我们提出了一种基于 DNA 的探针，称为“DNA 拉链”，它可以使用标准荧光显微镜在活细胞中对膜的有序性和瞬时相互作用的模式进行成像。通过精细调整 DNA 双链的长度和结合亲和力，这些探针可以通过动态 DNA 杂交精确延长膜脂相互作用的持续时间。我们还研究了膜有序性与 T 细胞受体信号激活之间的关系。这些可编程 DNA 探针在细胞孵育后即可发挥作用，它们可以很容易地适应不同的膜信号事件来研究脂质相互作用和膜有序性。
