Invertebrate model systems, such as Drosophila and Caenorhabditis elegans, provide an opportunity to identify novel genes that modulate acute responses to alcohol (Wolf and Heberlein, 2003). However, validation of these genes in mammalian systems, in which more complex aspects of alcoholrelated behaviors can be modeled, is necessary to provide relevance to human AUDs. Herein, we have used a technique to rapidly generate transgenic gene knockdown animals for behavioral testing. In theory, founder animals can be bred to maintain gene knockdown (Lois et al., 2002), although we found by breeding 1 of our founders that GFP fluorescence was maintained only through the first generation and appeared to be silenced in the second and third generations (data not shown). This may be due to silencing at the lentiviral integration site, perhaps due to toxicity from high shRNA expression (Cao et al., 2005; Grimm et al., 2006).
