Fluorophore-labelled RNA aptamers to common protein tags as super-resolution imaging reagents
Year: 2020
Type: preprint
Abstract: ABSTRACT Developing labelling methods that densely and specifically label targeted cellular structures is critically important for centroid localization-based super-resolution microscopy. Being easy and inexpensive to produce in the laboratory and of relatively small size, RNA aptamers have potential as a substitute for conventional antibody labelling. By using aptamers selected against common protein tags - GFP (green fluorescent protein) in this case - we demonstrate labelling methods using dSTORM-compatible fluorophores for STORM and hybridizable imager strands for DNA-PAINT super-resolution optical imaging of any cellular proteins fused to the aptamer binding target. We show that we can label both extracellular and intracellular proteins for super-resolution imaging, and that the method in particular, offers some interesting advantages for live cell super-resolution imaging of plasma membrane proteins. KEY POINTS A simple to use RNA aptamer method for super-resolution STORM and PAINT imaging in cells expressing common protein tags. The method has a number of unique advantages for live cell imaging at the nanometer scale. Provides a means to quantify the number of proteins being trafficked on the plasma membrane.
Institution: Cornell University
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: green
Funder: National Science Foundation
OpenAlex ID: W3006692895