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Engineering Yeast Extracellular Vesicle Biogenesis Through Rewiring Membrane Trafficking Pathways.

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This study enhances understanding of yeast-derived extracellular vesicle (EV) biogenesis using Saccharomyces cerevisiae. Researchers boosted EV production and cargo loading, revealing key protein sorting mechanisms for therapeutic applications.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Microbiology

Background:

  • Extracellular vesicles (EVs) show therapeutic potential, but their biogenesis mechanisms are not fully understood.
  • Saccharomyces cerevisiae is a safe, well-characterized yeast that naturally produces abundant EVs, making it a suitable model for study.
  • Elucidating EV biogenesis in yeast is crucial for harnessing their therapeutic capabilities.

Purpose of the Study:

  • To investigate the mechanisms of EV biogenesis and cargo loading in Saccharomyces cerevisiae.
  • To establish a tractable model system in S. cerevisiae for studying and engineering EV production.
  • To identify proteins involved in yeast-derived extracellular vesicles (YDEVs) biogenesis and cargo selection.

Main Methods:

  • Combined multicopy expression of chicken interferon-λ (ChiIFN-λ) with cell wall perturbation to increase EV yield.
  • Utilized quantitative proteomics to identify EV-associated proteins.
  • Performed functional analyses of key vesicle trafficking proteins (SNARE, ESCRT, Rab) and specific SNARE proteins (Sso2, Nyv1).

Main Results:

  • Achieved a tenfold increase in EV yield and efficient ChiIFN-λ incorporation into EVs.
  • Identified 1555 EV-associated proteins, including 501 transmembrane proteins from various organelles.
  • Demonstrated that ChiIFN-λ overexpression and cell wall stress alter vesicle trafficking regulators, reprogramming intracellular pathways.
  • Found Sso2 and Nyv1 SNARE proteins enriched in EV membranes, influencing EV size and subpopulations.

Conclusions:

  • Revealed conserved protein-sorting machinery governing yeast-derived extracellular vesicles (YDEVs) biogenesis.
  • Established Saccharomyces cerevisiae as a powerful platform for engineered EV production and mechanistic studies.
  • The findings provide insights into EV cargo loading and biogenesis for future therapeutic development.