Super-resolution biological microscopy would benefit from a much smaller alternative to green fluorescent protein and enzyme-mediated labelling methods for imaging specific proteins in cells. To facilitate routine tagging of organic fluorophores to cellular proteins for super-resolution imaging, we aim to combine genetic code expansion (the most non-invasive tagging method) with cell-compatible CuAAC with copper-chelating azides (the fastest bio-orthogonal reaction) and extend the utilities of the method to labelling of specific proteins in different subcellular locales, including intracellular.
The proposed work here is highly interdisciplinary, combining techniques from synthetic organic chemistry, in vitro evolution of novel protein function, optimizations of protein and chemical functions in mammalian cell context, and super-resolution protein imaging. The work is designed to take advantage of the fellow’s experience in protein labelling, bio-orthogonal chemistry and super-resolution imaging, along with the European host’s proficiency in in vitro evolution and genetic code expansion. The goal is to maximise the mutual transfer of knowledge while obtaining new methodologies and molecular insight into biological systems.
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