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Updated: Jun 20, 2026
08:54
In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
Published on: March 29, 2019
A mammalianized synthetic nitroreductase gene for high-level expression
Maik Grohmann1, Nils Paulmann, Sebastian Fleischhauer
1Department of Human Molecular Genetics, Max Planck Institute for Molecular Genetics, Berlin, Germany. grohmann@molgen.mpg.de
BMC Cancer
|August 29, 2009
Summary
Codon optimization of the nitroreductase gene improves its expression in mammalian cells. This enhances the nitroreductase/CB1954 system for cancer gene therapy by increasing prodrug activation and cell sensitivity.
Area of Science:
- Molecular Biology
- Biotechnology
- Cancer Gene Therapy
Background:
- The nitroreductase/CB1954 system is a promising gene-directed enzyme prodrug therapy (GDEPT) for cancer.
- It relies on bacterial nitroreductase (NTR) to activate the prodrug CB1954 into a cytotoxin.
- Improving NTR activity is key to enhancing this cancer treatment strategy.
Purpose of the Study:
- To enhance the therapeutic potential of the NTR/CB1954 system.
- To overcome expression limitations of bacterial nitroreductase in mammalian cells.
Main Methods:
- De novo synthesis of the bacterial nitroreductase gene with codon usage adapted for mammalian cells.
- Expression efficacy and sensitization to CB1954 were assessed using western blotting and cytotoxicity assays.
Main Results:
- A synthetic, codon-optimized nitroreductase gene (ntro) was generated with 144 silent base substitutions.
- ntro demonstrated significantly higher expression levels in mammalian cell lines compared to the native gene.
- Codon-optimized ntro increased cell sensitivity to CB1954 tenfold and improved the bystander effect.
Conclusions:
- Codon optimization effectively addresses translational expression barriers for bacterial genes in mammalian systems.
- This strategy significantly improves the NTR/CB1954 enzyme-prodrug system for cancer gene therapy.
- The enhanced NTR expression and activity pave the way for more effective human cancer treatments.
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