Invitrogen™ pSCREEN-iT™/lacZ-DEST Gateway™ Vector Kit

Description

Use the pSCREEN-iT™/lacZ-DEST Gateway™ Vector Kit to assess the potency of any RNAi knockdown reagent using a fast, sensitive, and cost-effective β-gal reporter assay or identify the best knockdown reagent for RNAi experiments without knowing protein function, western blotting, or qRT-PCR.
How it Works: The BLOCK-iT RNAi Target Screening system method uses a β-galactosidase activity assay to measure the knockdown ability of an RNAi reagent (Stealth RNAi™ siRNA, siRNA, dicer pools, or shRNA-containing plasmid). To identify the most effective knockdown reagent for your target, simply clone your target gene into the pSCREEN-iT /lacZ-DEST Gateway vector. Co-transfect the resulting construct with the knockdown reagent being tested. Expressed genes will be fused to β-galactosidase, enabling you to use β-galactosidase levels to measure the amount of RNAi-induced degradation of the target gene. The more effective any specific knockdown reagent is at mediating RNAi, the more degradation of the fusion mRNA will occur, lowering β-gal activity.
The BLOCK-iT RNAi Target Screening System includes the pSCREEN-iT /lacZ-DEST Gateway vector for generating a construct containing a lacZ/gene of interest fusion. This vector features:

• attR sites immediately following the lacZ ORF for fast and efficient recombination with any attL-flanked Gateway entry vector to generate the lacZ/target gene fusions
• The CMV promoter for strong expression of the lacZ/target gene fusion, leading to increased assay sensitivity and better discrimination between knockdown reagents of varied effectiveness
• Positional cloning to fuse a gene or fragment to the 3' end of the lacZ gene so that β-galactosidase will be expressed even if the target fragment being tested contains stop codons

RNAi, RNAi, Epigenetics and Non-Coding RNA Research, Vector-Based RNAi