Genetic code expansion (GCE)

We equip prion-propagating cell lines with a genetic code expansion (GCE) machinery for site-specific incorporation of an unnatural amino acid (UAA) into PrP. The UAA can be tagged with fluorescent dye suitable for super-resolution microscopy and cryo-CLEM/ET.

Our recently solved cryo-EM structures of authentic infectious ex vivo RML and ME7 prion fibrils combined with alanine scanning mutagenesis studies pinpointed candidate regions for the UAA insertion into PrP that supports prionogenic conversion.

The transgenic GCE cell platform will pave the way for the generation of a GCE mouse expressing UAA-PrP, which will enable direct observation of prion assemblies in mouse brain slices (in situ) by cryo-CLEM/ET.

Join us on this exciting journey!