TY - JOUR T1 - Procarcinogens - Determination and Evaluation by Yeast-Based Biosensor Transformed with Plasmids Incorporating RAD54 Reporter Construct and Cytochrome P450 Genes A1 - Ngoc Bui,Van A1 - Thu Huyen Nguyen,Thi A1 - Thanh Mai,Chi A1 - Bettarel,Yvan A1 - Yen Hoang,Thi A1 - Thuy Linh Trinh,Thi A1 - Hai Truong,Nam A1 - Ha Chu,Hoang A1 - Thanh Thanh Nguyen,Vu A1 - Duc Nguyen,Huu A1 - Woelfl,Stefan AD - VAST, Inst Biotechnol IBT, Natl Key Lab Gene Technol, Hanoi, Vietnam. AD - Heidelberg Univ, IPMB, Heidelberg, Germany. AD - Thai Nguyen Univ, Thai Nguyen Univ Sci, Thai Nguyen, Vietnam. AD - IRD, UMR MARBEC, Montpellier, France. AD - Vietnam Natl Univ Agr, Hanoi, Vietnam. UR - https://archimer.ifremer.fr/doc/00625/73740/ DO - 10.1371/journal.pone.0168721 N2 - In Vietnam, a great number of toxic substances, including carcinogens and procarcinogens, from industrial and agricultural activities, food production, and healthcare services are daily released into the environment. In the present study, we report the development of novel yeast-based biosensor systems to determine both genotoxic carcinogens and procarcinogens by cotransformation with two plasmids. One plasmid is carrying human CPR and CYP (CYP3A4, CYP2B6, or CYP2D6) genes, while the other contains the RAD54-GFP reporter construct. The three resulting coexpression systems bearing both CPR-CYP and RAD54-GFP expression cassettes were designated as CYP3A4/CYP2B6/CYP2D6 + RAD54 systems, respectively and used to detect and evaluate the genotoxic potential of carcinogens and procarcinogens by selective activation and induction of both CPR-CYP and RAD54-GFP expression cassettes in response to DNA damage. Procarcinogens were shown to be predominantly, moderately or not bioactivated by one of the CYP enzymes and thus selectively detected by the specific coexpression system. Aflatoxin B-1 and benzo(a) pyrene were predominantly detected by the CYP3A4 + RAD54 system, while N-nitrosodimethylamine only moderately activated the CYP2B6 + RAD54 reporter system and none of them was identified by the CYP2D6 + RAD54 system. In contrast, the genotoxic carcinogen, methyl methanesulfonate, was detected by all systems. Our yeast-reporter system can be performed in 384-well microplates to provide efficient genotoxicity testing to identify various carcinogenic compounds and reduce chemical consumption to about 53% as compared with existing 96-well genotoxicity bioassays. In association with a liquid handling robot, this platform enables rapid, cost-effective, and high-throughput screening of numerous analytes in a fully automated and continuous manner without the need for user interaction. Y1 - 2016/12 PB - Public Library Science JF - Plos One SN - 1932-6203 VL - 11 IS - 12 ID - 73740 ER -