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High Throughput Yeast Strain Phenotyping with Droplet-Based RNA Sequencing
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A high-throughput approach for studying virus replication in yeast.

Judit Pogany1, Tadas Panavas, Elena Serviene

  • 1University of Kentucky, Lexington, Kentucky, USA.

Current Protocols in Microbiology
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Yeast serves as a powerful model host for studying viral replication, enabling the identification of host factors essential for virus multiplication and functional interactions with viral components.

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Area of Science:

  • Microbiology
  • Virology
  • Molecular Biology

Background:

  • Viruses are obligate intracellular pathogens requiring host machinery for replication.
  • Identifying host factors is crucial for understanding viral life cycles and developing antivirals.
  • Model organisms offer advantages for dissecting complex host-pathogen interactions.

Purpose of the Study:

  • To highlight the utility of yeast as a model host for studying viral replication.
  • To describe high-throughput methods for analyzing host factors in viral multiplication.
  • To demonstrate the application of yeast systems for both plant and animal RNA virus research.

Main Methods:

  • Utilizing yeast's single gene-deletion and essential gene libraries (yTHC).
  • Controllable expression of viral proteins and nucleic acids in yeast.
  • Employing high-throughput screening procedures for host factor identification.

Main Results:

  • Yeast facilitates the identification of host factors critical for viral multiplication.
  • Functional interactions between host factors, viral proteins, and nucleic acids can be studied in yeast.
  • The described procedures are applicable to diverse RNA viruses, exemplified by tombusvirus (TBSV) and nodavirus (FHV).

Conclusions:

  • Yeast is a versatile and advantageous model system for virological research.
  • High-throughput yeast-based assays enable efficient discovery of host-virus interactions.
  • This approach advances the understanding of both plant and animal RNA virus replication mechanisms.