Application of SNAP-Tag in Expansion Super-Resolution Microscopy Using DNA Oligostrands

Frontiers in Chemistry

卷:9

Liwen Chen¹  Longfang Yao¹  Yiyan Fei¹  Lan Mi¹  Jiong Ma²  Li Zhang³ 

[1]Department of Optical Science and Engineering, Shanghai Engineering Research Center of Ultra-Precision Optical Manufacturing, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Green Photoelectron Platform, Fudan University, Shanghai, China

[2]|Insititute of Biomedical Engineering and Technology, Academy for Engineering and Technology, Fudan University, Shanghai, China

[3]|Shanghai Engineering Research Center of Industrial Microorganisms, The Multiscale Research Institute of Complex Systems, School of Life Sciences, Fudan University, Shanghai, China

关键词: expansion super-resolution;    DNA oligostrand;    SNAP;    F-actin;    nuclear pore complex;   

DOI  :  10.3389/fchem.2021.640519

来源: DOAJ

【 摘 要 】

Expansion super-resolution technology is a new technology developed in recent years. It anchors the dye on the hydrogel and the dye expands with the expansion of the hydrogel so that a super-resolution map can be obtained under an ordinary microscope. However, by labeling the target protein with a first antibody and secondary antibody, the distance between the fluorescent group and the actual target protein is greatly increased. Although fluorescent proteins can also be used for expansion super-resolution to reduce this effect, the fluorescent protein is often destroyed during sample preparation. To solve this problem, we developed a novel label system for expansion microscopy, based on a DNA oligostrand linked with a fluorescent dye, acrylamide group (linker), and benzoylguanine (BG, a small substrate molecule for SNAP-tag). This protocol greatly reduced the error between the position of fluorescent group and the actual target protein, and also reduced loss of the fluorescent group during sample preparation.

【 授权许可】

Unknown