Cardiovascular gene therapy aims to treat coronary and peripheral artery disease, heart failure and arrhythmia. Chosen transgene, delivery method, gene therapy vector type, high quality vector production and dose are all determining factors of the therapeutic outcome. High resolution vascular imaging and increased knowledge of vascular biology in physiological and pathological conditions enables finding of novel molecular targets for cardiovascular gene therapy. Transgenic and knockout mouse models have provided researchers several powerful experimental tools for studying the effects of single genes on cardiovascular diseases. For preclinical efficacy, safety and toxicology studies, large animal models are needed before entering to clinical testing. This review focuses on commonly used animal models in cardiovascular gene therapy and describes recent advancements in the field of vascular biology. Emphasis is also given for high resolution imaging of microvasculature and its impact on our knowledge of vascular function.