Genetically Encoded Multimeric (GEM) Tags for Intracellular Protein Localization in Cellular Cryo-ET

Le résumé fourni par la source

Cryo-ET now readily allows us to visualize the molecular sociology of cells in near-native state. However, cryo-ET is a label-free method — a major challenge remains in the precise localization of macromolecules of interest in the final tomographic volume. Solutions to this problem have included correlative light and electron microscopy (CLEM) and pattern recognition methods based on template matching or machine learning. However, these approaches can suffer from localization errors [1] or are easily applicable only to large biomolecules with identifiable structures [2]. Molecular tags in the form of genetically encoded multimeric particles (GEMs) offer a complementary visual solution for protein localization in tomograms. To develop an intracellular tag suitable for human cells, we conducted a library screen to identify icosahedron-forming proteins that can be functionalized for drug-inducible coupling to a target protein. Here we report on the performance of one GEM candidate, based on an archaeal encapsulin, in labelling endogenous proteins at different sub-cellular regions in human cells [3].

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