A high-throughput screen for genes essential for PRRSV infection using a piggyBac-based system

Area of Science:

• Virology
• Genetics
• Molecular Biology

Background:

• Porcine reproductive and respiratory syndrome virus (PRRSV) poses a significant threat to the swine industry.
• Understanding host-pathogen interactions is crucial for developing effective control strategies against PRRSV.

Purpose of the Study:

• To develop a high-throughput method for identifying host genes involved in PRRSV infection.
• To pinpoint specific genes that influence PRRSV susceptibility or resistance.

Main Methods:

• Utilized a dual-functional piggyBac transposon-based system for random mutagenesis in Marc145 cells.
• Challenged the mutated cell library with PRRSV and selected surviving clones.
• Employed inverse PCR and high-throughput sequencing to map transposon integration sites.
• Annotated flanking genes to identify candidate host factors.

Main Results:

• Generated a ranked list of candidate genes associated with PRRSV infection.
• Identified four high-priority genes (CDK17, RNF168, BCL2L15, TRIM33) significantly correlated with PRRSV.
• Validated the correlation of these genes in both Marc145 cells and porcine primary alveolar macrophages.

Conclusions:

• The study successfully identified key host genes influencing PRRSV infection.
• The piggyBac transposon system is a viable high-throughput tool for studying virus-host genetic interactions.
• Findings provide targets for future research into PRRSV pathogenesis and potential therapeutic interventions.