Rapad Universal Adenoviral Assay Kit

Background: Recombinant adenoviruses have tremendous potential in both research and therapeutic applications. There are numerous advantages they provide when introducing genetic material into host cells. The permissive host cell range is very wide. The virus has been used to infect many mammalian cell types (both replicative and non-replicative) for high expression of the recombinant protein. Recombinant adenoviruses are especially useful for gene transfer and protein expression in cell lines that have low transfection efficiency with liposome. After entering cells, the virus remains epichromosomal (i.e. does not integrate into the host chromosome so does not activate or inactivate host genes). Recently, recombinant adenoviruses have been used to deliver RNAi into cells. Two methods have traditionally been used to generate recombinant adenoviruses. The first involves homologous recombination of a shuttle vector containing gene of interest and an adenoviral backbone plasmid vector (restricted in E1/E3) in an adenovirus packaging cell line. The isolation of recombinant adenovirus by this method involves performing multiple plaque isolations to avoid wild-type virus and is extremely laborious and time consuming. The second method, pAdEasy system, employs the homologous recombination machinery in E. coli, a recombinant adenovirus is produced by a double-recombination event between cotransformed adenoviral backbone plasmid vector and a shuttle vector carrying the gene of interest. For the pAdEasy method, the system is high fidelity, but inefficient and requires the screening of many bacterial colonies. This results in a significant time commitment even before transfection of recombinant DNA into E1-expressing cells such as HEK293 cells. RAPAd Adenoviral Expression System provides a much faster and safer method to generate RCA-free recombinant adenovirus at high titer (see Table 1). The RAPAd system uses a novel Ad backbone devoid of the left-hand ITR, the packaging signal and E1 sequences. There is no need to perform the bacterial in vitro homologous recombination (pAdEasy method), and also the multiple plaque isolations (standard homologous recombination method in packaging cell line). The RAPAd system allows for generation of a recombinant virus within 2 weeks and the virus produced contained virtually no contaminating E1a sequences or replication-competent virus (RCA). RAPAd Adenoviral Expression System is simple to use. The method is straightforward and requires very limited 'hands on' time from shuttle/backbone cotransfection to the isolation of virus particles. It produces equivalent infectious titers as the standard viral genome/shuttle plasmid recombination method. In RAPAd Universal Adenoviral Expression System, the shuttle vector does not contain any promoter ahead of the multiple cloning sites. This allows you to introduce your own promoter that is optimal for your gene of interest or target cell. This makes the system ideal for promoter studies and cloning of shRNA.

Small volumes of RAPAd Universal Adenoviral assay kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS169396 is an assay kit targeting RAPAd Universal Adenoviral. Also known as RAPAd Universal Adenoviral Expression System.