Clonable In-situ Label for High-resolution Cellular Imaging
Cryo-electron tomography (cryo-ET) is a powerful technique that enables researchers to visualize macromolecular complexes inside cells at near-atomic resolution. However, localizing and identifying specific molecules in cryo-tomograms of cells remains a major challenge, as there is no green fluorescent protein (GFP)-like counterpart for molecular tagging and visualization in cryo-ET.
This group will develop a new generation of labels that are visible by cryo-ET of intact cells preserved in their native state. Through interdisciplinary collaborations, they will use a two-pronged approach of protein design and identification of naturally occurring proteins with specifically biased amino-acid composition in order to develop a widely-adaptable, cloneable label that generates a detectable gas bubble near the target protein when hit with a certain electron dose. Similar to the discovery of GFP, the technological development proposed here has the potential to transform structural proteomics and the field of cell biology as a whole.