Reshaping the yeast ER

Background.

The endoplasmic reticulum (ER) is the key organelle for folding and processing of secretory proteins in eukaryotes. Inefficient folding and secretion are a major challenge in production of recombinant proteins needed as biopharmaceuticals or for securing future food requirements. In most non-mammalian cells, the ER is spatially limiting these processes.

Aims & Methods.

The project SynthER - Designing a professional-secretory synthetic endoplasmic reticulum in yeasts and plant cells aims to redesign the internal membrane system and simultaneously enhance ER functionality in yeast and plant cells by mimicking the efficient ER structures of secretory mammalian cells.
Inspired by professional secretory mammalian cells, SynthER envisages to biomimic their stacked ER sheet and luminal architecture in yeast and plants. Dorian Stadlmayer will work on reshaping the ER in the yeast Komagataella phaffii (syn Pichia pastoris). The project will go beyond mere physical ER expansion by tailoring functionality through orthogonal and combinatorial expression of synthetic ER shaping proteins, using state-of-the art modular cloning techniques (e.g. Golden Gate cloning, CRISPR/Cas9-based genome engineering). The new morphologies will be monitored by a wide range of microscopic methods and quantitative image analyses, and their impact on the quality and quantity of secreted recombinant proteins will be determined.

Taken together, SynthER will develop novel cell factory concepts that represent a significant advance to the state of the art. Synthetic endomembrane engineering addresses a timely research topic through a pioneering synthetic biology approach with significant potential for advancing both the scientific understanding of secretory pathway plasticity and impacting technological applications, thus benefiting medicine and biotechnology. Our approach will leverage the strengths of interdisciplinary and cross-kingdom research, ensuring comprehensive analysis and optimization and facilitating the iterative design-build-test-learn cycle essential for refining synthetic ER structures.

Collaborations.

The work will be carried out in the framework of the WWTF-funded project “SynthER - Designing a professional-secretory synthetic endoplasmic reticulum in yeasts and plant cells“ at BOKU - Institute of Microbiology and Microbial Biotechnology, in close collaboration with Eva Stöger (BOKU - Institute of Plant Biotechnology & Cell Biology).