Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing

Area of Science:

• Biotechnology
• Molecular Biology
• Insect Cell Culture

Background:

• Adaptive laboratory evolution improves recombinant protein production in insect cells.
• Mechanisms behind enhanced influenza hemagglutinin (HA)-virus-like particle (VLP) expression in adapted cell lines are poorly understood.

Purpose of the Study:

• Investigate gene expression patterns in high-producer insect High Five cells adapted to neutral pH.
• Compare adapted cells to non-adapted cells during influenza HA-VLP expression.

Main Methods:

• RNA sequencing (RNA-seq) to analyze host cell and baculovirus gene expression.
• Differential gene expression analysis at 12 and 24 hours post-infection.
• Pathway analysis of differentially expressed genes.

Main Results:

• Adapted cells showed decreased host cell genome reads and increased baculovirus/transgene reads.
• 1742 host cell genes were differentially expressed; 474 at both time points.
• Differentially expressed baculovirus genes were down-regulated in adapted cells.
• Enriched pathways included ribosome biosynthesis, metabolism, oxidative phosphorylation, and protein processing.

Conclusions:

• Gene expression changes in adapted insect cells provide insights into mechanisms of enhanced HA-VLP production.
• Findings can guide engineering strategies for improved recombinant protein yields.
• Understanding host-pathogen interactions is key for optimizing VLP expression systems.