用于 RNA-蛋白质复合物共定位单分子光谱（CoSMoS）的 SNAP-Beads 的制备。

Preparation of SNAP-Beads for Colocalization Single-Molecule Spectroscopy (CoSMoS) of RNA-Protein Complexes.

机构信息

RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA.

出版信息

Methods Mol Biol. 2020;2113:17-22. doi: 10.1007/978-1-0716-0278-2_2.

PMID:32006304

Abstract

The SNAP-tag is a chemical tag that allows rapid and highly specific covalent labeling of proteins even in the full complexity of the cellular environment. The SNAP-tag has been instrumental to study native RNA-protein complexes at single-molecule resolution in their cellular environment as efficient labeling of the RNAs and proteins of interest is essential for this colocalization single-molecule spectroscopy (CoSMoS) technique. However, removal of excessive benzylguanine dye after the labeling reaction has remained challenging. Here, we describe a strategy to remove excessive benzylguanine dye using SNAP-tag coated beads as sponges.

摘要

SNAP 标签是一种化学标签，即使在细胞环境的完整复杂性中，也能允许快速且高度特异性的蛋白质共价标记。SNAP 标签对于在其细胞环境中以单分子分辨率研究天然 RNA-蛋白质复合物至关重要，因为有效标记感兴趣的 RNA 和蛋白质对于这种共定位单分子光谱学 (CoSMoS) 技术是必不可少的。然而，在标记反应后去除过量的苯甲基鸟嘌呤染料仍然具有挑战性。在这里，我们描述了一种使用 SNAP 标签包被珠作为海绵去除过量苯甲基鸟嘌呤染料的策略。