The collective attributes of adenoviruses (Ads), including ease of accomplishing replication deficiency, readily achievable high titers, encoding of large expression cassettes, efficiency of gene delivery to most cell types, and well-characterized biology, have made Ads, particularly Ad serotype 5 (Ad 5), some of the most utilized vectors for gene delivery. In recent years, however, it has become apparent that additional aspects of basic Ad virology must be uncovered for this vector system to succeed in the clinic. While local gene delivery is generally efficient, the broad tropism of Ad 5 and its tendency to home to the liver after systemic administration have proved to be limitations for other therapeutic applications, such as the treatment of disseminated cancers and cardiovascular disease. This has refocused research into the biology of Ad capsid components, particularly the main tropism determinant, the fiber/penton base complex, and their influence on transduction of selected cell types in vivo.