Reporter transgenes for study of oxidant stress in caenorhabditis elegans

For many studies of the effects of oxidant stress on cells, it can be advantageous to visualize the transcriptional response of the cell in vivo in real time. In optically transparent model systems, gene expression can be directly visualized by the construction of reporter transgenes expressing green fluorescent protein (GFP), as originally demonstrated by Chalfie and colleagues. This chapter describes both the general considerations involved in the construction of GFP reporter transgenes responsive to oxidative stress and the specific details of constructing a representative transgenic reporter in the model nematode worm Caenorhabditis elegans. Although the details of the representative reporter transgene apply specifically to C. elegans, the general approach should be applicable to many model systems. For genetic model systems such as C. elegans, oxidative stress-dependent induction of GFP expression represents a phenotype that can be the basis of forward genetic screens. These screens can potentially identify genes involved in the induction of oxidative stress response genes or genes that can be mutated to preemptively block oxidative stress in the first place.