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Regulation of exogenous gene expression by superoxide
Atsushi Kinoshita1, Daisuke Kobayashi, Yukiya Saitoh
1Department of Drug Informatics, Faculty of Pharmaceutical Sciences, Josai University, 1-1, Keyakidai, Sakado, Saitama, 350-0248, Japan.
Pharmaceutical Research
|October 19, 2006
Summary
This study shows that oxygen radicals can regulate gene expression from the cytomegalovirus long terminal repeat (CMV-LTR) promoter. This finding offers a new method for controlling exogenous gene expression in gene therapy vectors.
Area of Science:
- Molecular Biology
- Gene Therapy
- Biochemistry
Background:
- Gene expression regulation is crucial for gene therapy efficacy.
- The cytomegalovirus long terminal repeat (CMV-LTR) promoter contains a TPA response element (TRE) that binds activator protein 1 (AP-1).
- AP-1 is induced by oxidative stress, suggesting a potential regulatory mechanism.
Purpose of the Study:
- To investigate the regulation of exogenous gene expression driven by the CMV-LTR promoter using oxygen radicals.
- To explore the potential of oxidative stress as a tool for controlling gene expression in therapeutic vectors.
Main Methods:
- Two plasmids were used: pQBI25 (CMV-LTR with red-shift green fluorescent protein - rsGFP) and pRc/CMV-SOD (CMV-LTR with human Cu, Zn-superoxide dismutase - SOD).
- Cells were transfected and subsequently exposed to oxygen radicals generated by paraquat.
- Gene expression (c-fos, c-jun, rsGFP mRNA) and protein levels (rsGFP) were quantified using RT-PCR and fluorometry, respectively.
- Superoxide dismutase (SOD) activity was measured.
Main Results:
- Oxygen radical exposure significantly induced c-fos, c-jun, and rsGFP mRNA levels in FR-pQBI25 cells.
- A positive correlation was found between c-fos/c-jun mRNA and rsGFP mRNA levels.
- rsGFP protein levels were also significantly induced by oxygen radicals.
- Oxygen radical exposure significantly increased total SOD activity in L2-pRc/CMV-SOD cells.
Conclusions:
- Gene expression regulated by the CMV-LTR promoter can be modulated by oxygen radicals.
- Oxidative stress presents a viable strategy for controlling gene expression in CMV-LTR-based vectors.
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