Optimizing scaleup yield for protein production: Computationally Optimized DNA Assembly (CODA) and Translation...

Area of Science:

• Synthetic Biology
• Molecular Biology
• Biotechnology

Background:

• Altering gene translation kinetics can enhance protein production and function.
• Traditional gene synthesis methods may not optimize for translational efficiency.
• Codon context significantly impacts protein elongation rates and overall yield.

Purpose of the Study:

• To describe proprietary Translation Engineering technologies for designing synthetic genes.
• To present novel methodologies for improving protein structure, function, and production.
• To demonstrate the impact of codon pair usage and RNA secondary structures on translation.

Main Methods:

• Utilizing patented Translation Engineering technologies for gene design, accounting for codon pair usage, translational pausing, and RNA secondary structures.
• Employing Computationally Optimized DNA Assembly (CODA) for self-assembly of synthetic genes.
• Developing "Hot Rod" genes optimized for codon usage and devoid of translation-impeding elements.

Main Results:

• Engineered synthetic genes demonstrate high expression and enhanced protein production in various hosts, including cell-free systems.
• "Hot Rod" genes ensure maximal rates of coupled transcription and translation, protecting mRNA from degradation.
• Codon context, particularly over-represented codon pairs, influences protein folding and can be manipulated to solve expression challenges.

Conclusions:

• Translation Engineering combined with synthetic biology offers powerful tools to control gene expression and protein characteristics.
• Optimizing translation kinetics is crucial for overcoming limitations in heterologous protein expression, solubility, and activity.
• These patented technologies provide new avenues for restoring or enhancing protein function in diverse expression systems.