Invitrogen™ BLOCK-iT™ Adenoviral RNAi Expression System

Invitrogen™ BLOCK-iT™ Adenoviral RNAi Expression System

The pAd/BLOCK-iT™-DEST RNAi Gateway® Vector can be used for the efficient delivery and transient expression of a short hairpin RNA (shRNA) in vivo from an adenoviral vector. A novel cloning process places an ∼50-bp DNA oligonucleotide immediately following a U6 pol III promoter into the BLOCK-iT™ U6 entry vector. After efficient recombination of the entry vector into the pAd/BLOCK-iT™-DEST vector, followed by viral production and transduction, the shRNA driven by the U6 promoter can be transiently expressed in most dividing or non-dividing mammalian cell types. The shRNA generated avoids the hosts defense mechanism and will be effective at producing the RNAi gene knockdown response.

Features

• A promoterless region surrounded by attR sites for efficient recombination with the attL-flanked U6 Gateway® entry vector containing the RNAi cassette or any attL-flanked promoter and gene sequence
• All of the required components for efficient adenoviral packaging and delivery of the shRNA of interest. With this vector, transient analysis of gene knockdown in both dividing and non-dividing mammalian cell types and animal models can be achieved.

Contents

• The pAd/BLOCK-iT™ RNAi Gateway™ Vector is provided supercoiled at a concentration of 150 ng/µl and includes 10 µg of positive control vector. The BLOCK-iT™ RNAi Adenoviral Expression System includes the BLOCK-iT™ U6 Entry Vector Kit, the pAd/BLOCK-iT™-DEST vector and control, the 293A Cell line, and LR Clonase™ enzyme mix. Store vectors at -20°C. Store the cell line in liquid nitrogen. Store the LR Clonase™ mix at -80°C. Guaranteed stable for 6 months when properly stored.