5 – Adenoviral Vector Construction II: Bacterial Systems

Various methods have been developed to facilitate the generation of recombinant adenovirus vectors. Two commercially available methods employing bacterial systems have been most widely used: the homologous recombination method in bacteria and an in vitro ligation method based on simple routine plasmid construction. These methods can insert foreign genes not only into the E1 deletion region but also into the E3 deletion region, thereby allowing the construction of a binary transgene expression system in which heterologous genes can be inserted into both E1 and E3 regions. By modifying these methods, capsid-mutant adenovirus vectors and small-interfering RNA-expressing adenovirus vectors can be also constructed in order to control vector tropism and to induce the knockdown of gene expression, respectively. In this paper, we focus on the advances made so far in the methods for generating recombinant adenovirus vectors.