We summarize the current standard methods for overexpressing, inactivating, or manipulating genes, with special focus on nutritional and obesity research. These molecular biology procedures can be carried out with the maintenance of the genetic information to subsequent generations (transgenic technology) or devised to exclusively transfer the genetic material to a given target animal, which cannot be transmitted to the future progeny (gene therapy). On the other hand, the RNA interference (RNAi) approach allows for the creation of new experimental models by transient ablation of gene expression by degrading specific mRNA, which can be applied to assess different biological functions and mechanisms. The combination of these technologies contributes to the study of the function and regulation of different metabolism- and obesity-related genes as well as the identification of new pharmacologic targets for nutritional and therapeutic approaches.