Convenient Dual-Luciferase Reporter Assay Kit

The biological importance of non-coding DNA has recently emerged given the identification of single nucleotide polymorphisms (SNPs) in these regions. While regulatory regions that control gene expression such as promoters and enhancers are generally well defined, studies evaluating the alterations as result of SNPs in these regions remain limited. Importantly, luciferase assays are essential in delineating the function/activity of these regions and offer the ability to assess promoter and enhancer function in a robust manner. Here, I utilized the Dual-Luciferase reported kit from Promega to assess the impact of SNPs in promoter regions. 293T cells were transfected with pGL3-basic luciferase plasmids containing WT or mutant putative promoter regions upstream of the luciferase gene. These plasmids contain ~150bp non-coding DNA regions upstream of a luciferase gene. Transfection efficiency was measured by co-transfecting a Renilla luciferase plasmid with each pGL3-basic promoter variant. The Dual-Luciferase kit allows simultaneous readings of luciferase and renilla activity from the same sample, thus minimizing variation when reading samples.