Convenient Luminescence Signal Monitors RNAi Effect
The psiCHECK™-1 and psiCHECK™-2 Vectors are designed to provide quantitative and rapid optimization of RNA interference (RNAi). The vectors enable monitoring of changes in expression of a target gene fused to a reporter gene. In both vectors Renilla luciferase is used as the primary reporter gene, and the gene of interest is cloned into a multiple cloning region located downstream of the Renilla translational stop codon. Initiation of the RNAi process by synthetic siRNAs toward a gene of interest results in cleavage and subsequent degradation of the fusion mRNA. Measuring decreases in Renilla activity provides a convenient way of monitoring the RNAi effect.
In comparison with other fusion approaches (e.g., GFP or flag-tags), the Renilla luciferase approach offers more convenient and rapid quantitation with higher sensitivity. The psiCHECK™-1 Vector is recommended for use in monitoring RNAi effects in live cells. The changes in Renilla luciferase activity are measured with the EnduRen™ Live Cell Substrate (Cat.# E6481), which allows continuous monitoring of intracellular Renilla luminescence. The psiCHECK™-2 Vector contains a second reporter gene, firefly luciferase, and is designed for endpoint lytic assays. Introduction of firefly luciferase in the psiCHECK™-2 Vector allows normalization of Renilla luciferase expression, achieving robust and reproducible results.
psiCHECK™ Vector Advantages for RNAi Optimization
- Convenient RNAi target evaluation.
- Quantitation is performed with a common luminometer; no need to purchase expensive equipmenton.
- Protocols allow for measurements in live cells or crude cell lysates.
- No labor-intensive, time-consuming assays or waiting for phenotypic changes.
- No need for transfection normalization when using the psiCHECK™-2 Vector.
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For Research Use Only. Not for Use in Diagnostic Procedures.