Cyagen provides several solutions:
●Fusion tag technology: Proprietary hydrophilic fusion partners (e.g., YaiN/YbeL analogs) enhance solubility and purification efficiency for membrane proteins, achieving >80% success rates in functional reconstitution
●PTM-aware design: AI-driven sequence optimization identifies problematic PTM sites preemptively, enabling codon-optimized synthetic genes that bypass aggregation-prone regions while preserving functional domains
●Hybrid expression systems: Targeted-gene-editing-engineered HEK293 lines with inducible secretory pathway boosters enable authentic glycosylation and phosphorylation for therapeutic proteins

The primary challenges include:
●Membrane protein insolubility: Hydrophobic regions often lead to aggregation, particularly in E. coli systems
●PTM incompatibility: Prokaryotic systems lack native eukaryotic post-translational modification machinery, risking misfolding and loss of functionality
●Cellular toxicity: Overburdened secretion pathways can impair cell viability during high-yield expression

Cyagen implements several strategies:
●Inducible systems: Tetracycline- or ligand-regulated promoters allow controlled expression timing, reducing prolonged stress.
●Optimized delivery: Our 3rd-generation lentiviral systems achieve high transduction efficiency at lower viral titers, minimizing off-target effects.
●Promoter selection: Strong but regulated promoters like CAG enable high expression without requiring excessive plasmid DNA, lowering metabolic burden.

We employ advanced multiparametric codon optimization algorithms that adjust codon bias to match the host cell’s translational machinery while stabilizing mRNA secondary structures. This approach, validated by industry-standard tools, increases protein yield without altering the amino acid sequence. For example, codon-optimized genes in HEK293 cells have demonstrated up to 86% improved expression in functional studies.

Cyagen utilizes the CAG promoter (CMV enhancer fused to the chicken beta-actin promoter), which demonstrates superior performance in maintaining high-level, stable transgene expression compared to traditional promoters like CMV. This promoter drives stronger mRNA and protein yields in HEK293F cells, even under suboptimal conditions. For lentiviral-based systems, Cyagen’s targeted gene editing platform incorporates optimized promoters tailored to your specific cell line, ensuring broad tropism and minimal transcriptional silencing.