psiCHECK™-1 and psiCHECK™-2 Vectors

Vectors for Optimizing RNA Interference Assays

• Monitor changes in expression of a target gene fused to a reporter gene; decreases in luciferase activity correlate with RNAi
• psiCHECK™-1 Vector is used to monitor RNAi effects in live cells
• psiCHECK™-2 Vector designed for endpoint lytic assays

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.

ItemPart #Size

psiCHECK™-2 Vector

C802A1 × 20μg

แหล่งที่มา : https://worldwide.promega.com/products/rna-analysis/rna-interference/psicheck-1-and-psicheck-2-vectors/?catNum=C8021