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Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

Department of Marine Biology, Ocean University of China, Qingdao 266003, China
Author to whom correspondence should be addressed.
Biosensors 2012, 2(3), 318-340;
Received: 27 July 2012 / Revised: 30 August 2012 / Accepted: 6 September 2012 / Published: 10 September 2012
(This article belongs to the Special Issue Nano and Micro DNA/RNA Sensors)
PDF [339 KB, uploaded 10 September 2012]


p21CIP1/WAF1 is a p53-target gene in response to cellular DNA damage. Here we report the development of a fish cell biosensor system for high throughput genotoxicity detection of new drugs, by stably integrating two reporter plasmids of pGL3-p21-luc (human p21 promoter linked to firefly luciferase) and pRL-CMV-luc (CMV promoter linked to Renilla luciferase) into marine flatfish flounder gill (FG) cells, referred to as p21FGLuc. Initial validation of this genotoxicity biosensor system showed that p21FGLuc cells had a wild-type p53 signaling pathway and responded positively to the challenge of both directly acting genotoxic agents (bleomycin and mitomycin C) and indirectly acting genotoxic agents (cyclophosphamide with metabolic activation), but negatively to cyclophosphamide without metabolic activation and the non-genotoxic agents ethanol and D-mannitol, thus confirming a high specificity and sensitivity, fast and stable response to genotoxic agents for this easily maintained fish cell biosensor system. This system was especially useful in the genotoxicity detection of Di(2-ethylhexyl) phthalate (DEHP), a rodent carcinogen, but negatively reported in most non-mammalian in vitro mutation assays, by providing a strong indication of genotoxicity for DEHP. A limitation for this biosensor system was that it might give false positive results in response to sodium butyrate and any other agents, which can trans-activate the p21 gene in a p53-independent manner. View Full-Text
Keywords: genotoxicity; fish cells; p21; p53; trans-activation; luciferase genotoxicity; fish cells; p21; p53; trans-activation; luciferase

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Geng, D.; Zhang, Z.; Guo, H. Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression. Biosensors 2012, 2, 318-340.

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