This RNA intereference assay is designed to monitor knockdown of RNAi target sequences efficiently and easily, using a no-cell-lysis protocol. It consists of the pRNAiMonitor-MetLuc2 Vector and the Ready-To-Glow Secreted Luciferase Reporter Assay. The pRNAiMonitor-MetLuc2 vector expresses MetLuc, a secreted luciferase reporter that can easily be detected in the medium surrounding the cells. When a target sequence (encoding the target of your shRNA or siRNA) is cloned downstream of the MetLuc gene, a bicistronic mRNA transcript encoding both the MetLuc and the target sequence is produced. Target gene knockdown results in the degradation (or translational inhibition) of the mRNA transcript, which causes a proportional decrease in the expression of the reporter protein. The reporter molecule is easily detected with the Ready-To-Glow Secreted Luciferase Reporter Assay kits.