Adeno-X Adenovirus Vector System 3

Features & Benefits

• Fastest, easiest adenovirus vector system ever
• Clone directly into pAdenoX vector without shuttle vector
• >90% cloning efficiency in just 2–3 days
• Very high titer, high expression, broad host range

Description

Until now the main drawback of commercially supplied adenoviral vectors has been the need to use complex cloning procedures to overcome the difficulties with cloning into large (~34 kb) plasmids. Clontech adenovirus virologists have harnessed the power of In-Fusion HD PCR cloning technology to create the easiest adenoviral cloning method.

In the Adeno-X 3 system DNA sequences can be rapidly transferred as PCR products to any pAdenoX vector using the In-Fusion cloning method. Your gene of interest is amplified with 15 bp extensions that are homologous to the ends of the linearized adenoviral vector. The PCR product is then purified and mixed with the linearized adenoviral vector of choice in the In-Fusion reaction. Following the reaction, a portion of the mixture is transformed into Stellar Competent Cells and screened. Once a PCR-positive clone is identified, the recombinant adenoviral vector is amplified, purified, and subsequently linearized with the restriction enzyme PacI, then transfected into HEK 293 cells for viral rescue and amplification.

Multiple Formats Are Available
Adeno-X Adenoviral System 3 is available in seven formats, including the most advanced tetracycline inducible expression system, constitutive expression systems with or without fluorescent reporters, and universal ones that allow you to clone and express any entire cassette of your choice.

When you clone your gene into pAdenoX-Tet3G you are creating a system with the tightest and most sensitive control of gene expression. Tightly-controlled, doxycycline-induced expression is as easy as constitutive expression since the Tet-On 3G transactivator protein and the PTRE3G controlled gene of interest are contained on the same adenoviral vector. Up to 3000-fold induction can be archived using this system (see Images & Data tab).

Clone Any Expression Cassette Into the Universal Vectors
You are not limited to using a CMV expression system—we have created universal systems with vectors that lack a promoter and polyA signal in the cloning site. Simply amplify an entire expression cassette (from promoter to polyA) from a pre-existing construct and clone using In-Fusion HD. Universal systems can be used for expression from alternative promoters that are more suitable to your target cell type such as EF1 alpha or tissue-specific promoters. Alternatively, you may wish to transfer your shRNA or miRNA expression cassette from a pre-existing plasmid to one of the universal pAdenoX vectors in order to create a high efficiency RNAi delivery system. Even if your expression cassette does not exist you can create one using multiple fragment cloning.