Codon Optimized Factor IX Gene Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current treatments for hemophilia B, such as plasma-derived Factor IX and recombinant Factor IX, suffer from challenges like low concentration in plasma, difficulty in purification, need for frequent dosing due to short half-life, and inefficiency in heterologous expression systems.
Innovation Solution
The development of an isolated nucleic acid molecule with a nucleotide sequence at least 85% identical to SEQ ID NO:1, which encodes a polypeptide with Factor IX activity, optimized for enhanced expression and stability, including codon optimization, increased G/C content, and incorporation of heterologous sequences for improved half-life and pharmacokinetic properties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If plasma-derived Factor IX is purified from plasma, then active Factor IX is obtained, but the purification process is very difficult due to low concentration in plasma
Solution Approach 1:
The patent uses recombinant DNA technology to create a copy of the Factor IX gene and express it in heterologous host cells (such as CHO cells, HEK293 cells, or NS0 cells). This produces recombinant Factor IX (rFIX) that mimics the natural protein without requiring purification from human plasma, thereby solving the contradiction between obtaining active Factor IX and the difficulty of purification.
2Reliability
If plasma-derived Factor IX is used for treatment, then Factor IX replacement is achieved, but frequent dosing is required due to short half-life
Solution Approach 1:
The patent employs codon optimization to modify the nucleotide sequence of the Factor IX gene, increasing the G/C content and using codons preferred by the host cell's translational machinery. This optimization enhances the stability and half-life of the recombinant Factor IX protein, reducing the frequency of dosing required while maintaining therapeutic efficacy.
3Productivity
If recombinant Factor IX is expressed in heterologous systems, then production is achieved, but expression efficiency is low
Solution Approach 1:
The patent optimizes the nucleotide sequence by increasing G/C content and selecting codons that are preferentially used by the host cell's tRNA pool. This codon optimization dramatically improves translation efficiency and protein expression levels in heterologous systems, resolving the contradiction between achieving production and maintaining ease of manufacture.
4Productivity
If the nucleotide sequence is optimized with increased G/C content and codon optimization, then expression and stability are enhanced, but the sequence diverges from the native sequence
Solution Approach 1:
The patent strategically modifies the nucleotide sequence by optimizing codon usage and increasing G/C content in regions that do not affect the amino acid sequence or critical functional domains. This approach enhances expression and stability while preserving the essential biological function and structure of Factor IX, thereby balancing productivity gains with compositional stability.
Data Source
AI summary
The present invention provides codon optimized Factor IX sequences, vectors and host cells comprising codon optimized Factor IX sequences, polypeptides encoded by codon optimized Factor IX sequences, and methods of producing such polypeptides. The present invention also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject a codon optimized Factor IX nucleic acid sequence or the polypeptide encoded thereby.