规格
Cloning Method:	Gateway®
Constitutive or Inducible System:	Constitutive
Delivery Type:	Adenoviral
Product Line:	BLOCK-iT™
Promoter:	U6
RNAi Type:	shRNA
Vector:	pAd

储存

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.

描述

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.

The pAd/BLOCK-iT™-DEST vector (Figure 1) offers:

• 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.

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