Construcion of a Helper-dependent adenovirus carrying gene coding for human Interleukin 12

The clinical success of gene therapy depends on the development of suitable gene transfer vectors. High efficiency gene delivery, a favorable safety property, and feasibility of large-scale production are the most important. Helper-dependent adenovirus vector (HDAd) with it deleted of all viral coding sequences, have a higher cloning capacity, improved performance of tissue specific promoter, and reduced toxicity in animals making these vectors promising tools for gene transfer in vitro and in vivo . However, HDAd vectors are very difficult to work with than standard expression plasmids because it is the large size plasmids with a few unique restriction enzyme sites so that it made difficult to clone transgene insertion into that vector. Gateway Technology provides the ideal route to the ViraPower viruses vector expression system. There are several ViraPower expression systems for Lentiviral and Adenoviral are now available from Invitrogen. However, a ViraPower expression system for Helper-dependent adenovirus is not yet develops. We work with Helper-dependent adenovirus vector by cloning DNA fragment sequences that flank by attR sites to generation of the HdAd-gfp-DEST expression vector to create the new highthroughput Helper-dependent adenovirus Destination vector. Latter, this vector system is use to make a virus recombinant expressing human Interleukin-12.