Beta-Ca²⁺/calmodulin-dependent protein kinase II (βCaMKII) is known as a powerful regulator of lateral habenula neuron function and a key molecular determinant of depression. βCaMKII is also abundant in the hippocampus, especially in the dentate gyrus (DG). However, whether βCaMKII in the DG is also involved in emotional behaviors remains unknown. In this study, using βCaMKII-F90G transgenic mice, in which the overexpression of βCaMKII is restricted to the DG, we demonstrated that βCaMKII overexpression in the DG has no effect on the level of anxiety in mice in open field test or light–dark box test. Moreover, tail suspension test and forced swim test showed that the level of depression in βCaMKII-F90G transgenic mice and their littermates was comparable. Taken together, our findings indicate that βCaMKII overexpression in the DG does not result in depression- and anxiety-like behaviors in mice and provide evidence that the function of βCaMKII in emotional behaviors is brain region specific.