Invitrogen™ BLOCK-iT™ Lentiviral RNAi Gateway™ Vector Kit

Description

A novel cloning process places an approx. 50-bp DNA oligonucleotide immediately following a U6 pol III promoter into the BLOCK-iT U6 entry vector. The oligonucleotide is designed to express RNA that forms a stemLoop structure containing the sense and antisense regions of your target gene of interest. This shRNA is then recombined into the pLenti6/BLOCK-iT-DEST vector. After viral production and transduction, the shRNA driven by the U6 promoter becomes stably integrated as an RNAi cassette. The shRNA generated avoids the hosts defense mechanism and will be effective at producing the RNAi gene knockdown response. Using the BLOCK-iT Lentiviral RNAi Expression System, long-term analysis of gene blocking in both dividing and non-dividing mammalian cell types and animal models can be achieved.

The BLOCK-iT RNAi U6 Entry Vector Kit allows streamlined cloning of shRNA target sequences for testing in transient experiments. Selected RNAi expression cassettes are quickly and efficiently recombined from the BLOCK-iT RNAi U6 entry vector into the pLenti6/BLOCK-iT-DEST vector via a standard Gateway LR recombination reaction.

• attR sites for efficient recombination with the attL-flanked U6 Gateway™ entry vector containing the RNAi cassette
• All of the required components for efficient lentiviral packaging and delivery of the shRNA of interest
• Blasticidin selection marker for fast, efficient selection of stable cell lines expressing the shRNA

Cloning, Gateway Cloning, Lentiviral shRNA and miRNA, RNAi, RNAi, Epigenetics and Non-Coding RNA Research, Virus-Based RNAi