Related Experiment Video
Updated: Jun 13, 2025
14:09
Genetic Studies of Human DNA Repair Proteins Using Yeast as a Model System
Published on: March 18, 2010
18.1K
A Humanized Yeast Model for Studying TRAPP Complex Mutations; Proof-of-Concept Using Variants from an Individual with
Erta Zykaj1, Chelsea Abboud1, Paria Asadi1
1Department of Biology, Concordia University, Montreal, QC H4B1R6, Canada.
Cells
|September 14, 2024
Summary
Humanized yeast models using CRISPR/Cas9 technology efficiently study genetic variants in TRAPP subunits, aiding diagnosis of rare membrane trafficking disorders and neurodevelopmental conditions.
Area of Science:
- Cell Biology
- Genetics
- Molecular Biology
Background:
- Genetic variants in membrane trafficking proteins, including TRAPP complexes, cause severe rare disorders.
- TRAPP complexes regulate membrane trafficking and autophagy, and variants are linked to neurological and muscular diseases.
- Studying these variants is crucial for diagnosis and understanding TRAPP subunit functions, but patient cells are often unavailable.
Purpose of the Study:
- To develop and validate a humanized yeast model for studying TRAPP subunit variants.
- To investigate the functional impact of TRAPPC1 variants associated with a severe neurodevelopmental disorder and myopathy.
- To demonstrate the utility of humanized yeast for disease modeling and variant interpretation.
Main Methods:
- Humanized yeast models were created using CRISPR/Cas9 to replace core yeast TRAPP subunits with human orthologs.
- Specific TRAPP subunits (TRAPPC1, TRAPPC2, TRAPPC2L, TRAPPC6A, TRAPPC6B) were successfully humanized.
- The humanized yeast system was used to analyze pathogenic TRAPPC1 variants from a patient with neurodevelopmental disorder and myopathy.
Main Results:
- Human TRAPPC1, TRAPPC2, TRAPPC2L, TRAPPC6A, and TRAPPC6B successfully replaced their yeast counterparts.
- Analysis of TRAPPC1 variants revealed a non-functional maternal variant and a conditional-lethal paternal variant affecting secretion and autophagy.
- Observed defects in humanized yeast paralleled those in patient fibroblasts and were rescued by wild-type TRAPPC1.
Conclusions:
- Humanized yeast models provide an efficient platform for studying TRAPP subunit variants when patient cells are scarce.
- This system aids in assigning clinical significance to variants of unknown significance (VUS) and facilitates disease diagnosis.
- The study establishes a foundation for characterizing additional TRAPP variants using this humanized yeast model.
