Enhanced expression of codon optimized interferon gamma in CHO cells.

Area of Science:

• Biotechnology
• Molecular Biology
• Protein Expression

Background:

• Human interferon-gamma (IFN-γ) is a promising therapeutic agent with immunomodulatory functions.
• Chinese hamster ovary (CHO) cells are widely used for industrial-scale recombinant protein production.
• Suboptimal foreign protein expression in CHO cells can result from non-native codon patterns.

Purpose of the Study:

• To apply a novel codon optimization approach for designing synthetic IFN-γ coding sequences.
• To enhance heterologous expression of IFN-γ in CHO cells.
• To compare the effectiveness of individual codon usage (ICU) and codon context (CC) optimization strategies.

Main Methods:

• RNA-Seq based transcriptome profiling of CHO cells to determine gene expression patterns.
• Design of synthetic IFN-γ genes optimized for ICU and CC.
• In vivo expression analysis of wild-type and codon-optimized IFN-γ in CHO cells.

Main Results:

• CHO cell transcriptome analysis revealed similar ICU and CC distribution patterns across different gene expression levels.
• Codon context (CC) optimized IFN-γ demonstrated a significant >13-fold increase in expression compared to wild-type.
• Individual codon usage (ICU) optimized IFN-γ showed a maximum 10-fold increase in expression.

Conclusions:

• Codon context is a more effective parameter than individual codon usage for improving recombinant IFN-γ expression in CHO cells.
• The findings provide a refined strategy for optimizing heterologous gene expression in mammalian cell hosts.
• This work advances the efficient production of therapeutic proteins like IFN-γ.