Codon-Optimized Factor VIII Nucleic Acids for Expression
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Solution Overview
Problem
The high cost of commercial production and poor expression of recombinant Factor VIII protein in heterologous expression systems due to cis-acting elements, such as transcriptional silencers and matrix attachment-like sequences, hinder the development of a low-cost recombinant FVIII protein for patients.
Innovation Solution
Codon-optimized nucleic acid molecules encoding polypeptides with Factor VIII activity, including genetic cassettes with specific promoters, enhancers, and sequences that enhance expression, are used to improve the yield and therapeutic efficacy of FVIII protein production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the native FVIII coding sequence is used in heterologous expression systems, then the protein can be produced, but the expression level is two to three orders of magnitude lower than similarly sized proteins due to cis-acting elements
Solution Approach 1:
The patent removes harmful cis-acting elements (transcriptional silencer elements, matrix attachment-like sequences, and transcriptional elongation inhibitory elements) from the native FVIII coding sequence. This extraction of detrimental sequences allows the remaining FVIII gene to be expressed at much higher levels in heterologous systems without the suppressing effects of these elements.
Solution Approach 2:
The patent modifies the coding sequence parameters of FVIII by altering nucleotide sequences while maintaining amino acid sequence integrity. This includes optimizing codon usage and removing inhibitory cis-acting elements, which changes the transcriptional and translational parameters to achieve higher expression levels in heterologous expression systems.
2Reliability
If recombinant FVIII protein is produced commercially, then patients can receive treatment, but the production cost remains high due to poor expression yields
Solution Approach 1:
The patent optimizes the nucleotide sequence parameters of FVIII to improve expression efficiency in heterologous systems. By modifying codon usage and removing inhibitory elements, the production yield increases significantly, which directly reduces the cost per unit of therapeutic protein while maintaining reliable supply for patient treatment.
Data Source
AI summary
The present disclosure provides codon optimized Factor VIII sequences, vectors, and host cells comprising codon optimized Factor VIII sequences, polypeptides encoded by codon optimized Factor VIII sequences, and methods of producing such polypeptides.