Major attention has been focused on the development of gene therapy approaches for the treatment of vascular diseases. In this review, we focus on an alternative use of gene therapy: the use of genetic means to study vascular cell biology and physiology. Both viral and nonviral gene transfer strategies have limitations, but because of the overwhelming inflammatory responses associated with the use of viral vectors, nonviral gene transfer methods are likely to be used more abundantly for future applications in the vasculature. Researchers have made great strides in the advancement of gene delivery to the vasculature in vivo. However, the efficiency of gene transfer seen with most nonviral approaches has been exceedingly low. We discuss how to circumvent and take advantage of a number of the barriers that limit efficient gene delivery to the vasculature to achieve high-level gene expression in appropriate cell types within the vessel wall. With such levels of expression, gene transfer offers the ability to alter pathways at the molecular level by genetically modulating the activity of a gene product, thus obviating the need to rely on pharmacological agents and their foreseen and unforeseen side effects. This genetic ability to alter distinct gene products within a signaling or biosynthetic pathway or to alter structural interactions within and between cells is extremely useful and technologically possible today. Hopefully, with the availability of these tools, new advances in cardiovascular physiology will emerge.