Development of a cellular biosensor system for genotoxicity detection based on Trp53 promoter

Area of Science:

• Biotechnology
• Toxicology
• Molecular Biology

Background:

• Genotoxicity testing is crucial for assessing chemical safety.
• Existing methods can be time-consuming and lack high-throughput capabilities.
• Development of sensitive and specific biosensors is needed for efficient chemical screening.

Purpose of the Study:

• To create a mouse cell biosensor for high-throughput genotoxicity detection.
• To enable rapid identification of environmental pollutants and new drug candidates.
• To establish a sensitive and specific in vitro system for genotoxicity assessment.

Main Methods:

• A novel reporter vector, pGL4-p53-GFP, was constructed, linking the p53 promoter to the green fluorescent protein (GFP) gene.
• This reporter was stably integrated into mouse embryonic fibroblast cells to create a biosensor system.
• The system's response to genotoxic agents was evaluated via fluorescence microscopy and Western blot analysis.

Main Results:

• The biosensor demonstrated significantly increased GFP expression upon exposure to genotoxic agents like doxorubicin, cyclophosphamide, and benzo(a)pyrene.
• The system showed a negative response to the non-genotoxic agent dimethyl sulfoxide, confirming its specificity.
• These results highlight the biosensor's sensitivity and specificity in detecting genotoxicity.

Conclusions:

• The developed mouse cell biosensor system offers a novel and effective in vitro approach for genotoxicity detection.
• This system is particularly useful for high-throughput screening of chemicals, including environmental pollutants and pharmaceuticals.
• The biosensor provides a sensitive and specific tool for advancing genotoxicity assessment.