Codon-Optimized Sodium-Iodide Symporter Gene for Eukaryotic Expression
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Solution Overview
Problem
Current modified Sodium-Iodide Symporter (NIS) genes are not optimized for efficient expression in eukaryotic cells, particularly human cells, which hampers their application in thyroid disease treatment and radionuclide gene therapy and molecular imaging.
Innovation Solution
A codon-optimized polynucleotide encoding the Sodium-Iodide Symporter (NIS) protein is developed, specifically modifying codons in the 8th transmembrane domain, 5th transmembrane domain, C-terminal region, phosphorylation sites, and N-glycosylation sites to enhance expression in eukaryotic cells, including human cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the original NIS gene sequence is used for expression in eukaryotic cells, then the amino acid sequence is preserved, but the expression efficiency is insufficient
Solution Approach 1:
The patent applies parameter changes by modifying nucleotide codons while preserving the amino acid sequence. Specifically, it optimizes codon usage frequency to match eukaryotic cell preferences, adjusts GC content to improve transcriptional efficiency, and modifies codon positions in transmembrane domains to enhance protein folding and stability. These parameter changes resolve the contradiction by improving expression efficiency without altering the functional amino acid sequence of the NIS protein.
2Productivity
If codon optimization is performed to enhance expression in eukaryotic cells, then expression efficiency improves, but the complexity of gene modification increases
Solution Approach 1:
The patent applies local quality by selectively optimizing specific regions of the NIS gene rather than uniformly modifying the entire sequence. It focuses codon optimization on transmembrane domains (5th, 8th, and 12th domains) and the C-terminal region, which are critical for protein stability and function. This localized approach enhances expression efficiency while minimizing the overall complexity of gene modification compared to comprehensive sequence redesign.
Data Source
AI summary
The present disclosure provides a polynucleotide which is codon optimized for the efficient expression in a eukaryotic cell, a plasmid and a eukaryotic cell comprising the same. The modification resulted in the efficient expression of NIS in eukaryotic cells and the enhancement of the function of NIS by glycosylation. Thus modified polynucleotide encoding NIS of the present disclosure is useful as imaging reporter for gene, viral and/or cell based therapies.