Click chemistry-based high speed labelling reactions for super resolution microscopy
Advances in light microscopy have honed technologies such as optics, instrumentation and software. However, there is still a gap in development of better labelling techniques. The EU-funded NUPTAG (Protein-engineering based approach for super-resolution imaging of nucleoporin-chromatin interactions) project has therefore developed new protein labelling methods with small dye molecules. Combining state-of-the-art chemical biology and protein engineering has produced the synergy to achieve this. Researchers used genetic code expansion and a unique combination of two very fast chemical reactions for successful dual colour protein labelling. The methodology has been published in several journal articles and is the basis for a patent application. The new technique was able to label distinct populations of membrane proteins such as insulin receptors, and viral like particles. Further research goals focus on applications to study nuclear pores, which act as channels for transport of materials and proteins in and out of the nucleus to control e.g. gene expression. NUPTAG work has opened up many possibilities for non-invasively labelling cells. The tag is likely to be the smallest available at the moment and the protocol can be applied to living cells, a very significant advantage. The research team is active in collaborations worldwide and is still refining the techniques.
Keywords
Click chemistry, super-resolution microscopy, fluorescent dye, NUPTAG, nuclear membrane, genetic code expansion