Sensitive DsRed fluorescence-based reporter cell systems for genotoxicity and oxidative stress assessment.

摘要: Various in vitro test systems have been developed for genotoxic risk assessment early drug development. However, these genotoxicity tests often show limited specificity, and provide insights into the mode of toxicity tested compounds. To identify genes that could serve as specific biomarkers or oxidative stress, we exposed mouse embryonic stem (ES) cells to various stress-inducing compounds performed genome-wide expression profiling. Differentially expressed were classified based on fold-change their specificity either stress. Promoter regions four selected (Ephx1, Btg2, Cbr3 Perp) fused a DsRed fluorescent reporter gene stably integrated ES cells. Established stable cell lines displayed significant induction upon exposure different classes In contrast, non-genotoxic carcinogenic did not induce even at cytotoxic doses. Expression Cbr3-DsRed was more responsive stress while other three reporters reacted direct-acting agents. Therefore, differential response Btg2- can indicator main action mechanism addition, evidence inhibition DNA replication results preferential activation Btg2-DsRed reporter. conclusion, generated sensitive allow detection properties chemical be used high-throughput assays.