Biotic resistance is often posited, but rarely known, to be the cause of invasion failure. Competition and predation are the most frequently identified processes that may prevent or limit the establishment of nonnative species. Interactions between nonnative and native species that involve intraguild predation (IGP) are very common in nature, although theory predicts most IGP systems should be unstable and lead to extinction. If this prediction is true, the frequency of invasion failures due to IGP may be underappreciated because of their fleeting nature and, thus, studies of unstable IGP systems are lacking, despite the opportunities they offer for understanding the factors affecting their unstable dynamics. We investigated a failed invasion involving an IGP relationship. In Florida, the guppy (Poecilia reticulata), a worldwide invader, fails to establish in the presence of eastern mosquitofish (Gambusia holbrooki). We tested whether and how resident mosquitofish cause guppy invasion failure using replicated mesocosm and aquarium trials. Both the predator and competitor components of the IGP relationship were strongly asymmetrical, with large impacts on guppies. We identified two effects, direct consumption of neonates and aggressive interference competition, that limited survival and recruitment. The highly unstable nature of this IGP relationship is the primary cause of the failure of the guppy to establish in Florida. Our study shows that the transient nature of an ephemeral IGP relationship can yield important insights into the underlying causes of invasion failure, including the role of strong biotic resistance.