Understanding the molecular mechanisms underlying anxiety and ethanol-related behaviors is crucial for developing effective therapeutic interventions. This study identifies a novel role for microRNA miR-181a and its target, Sirtuin 1 (SIRT-1), in the hippocampus as contributors to anxiety-like behavior and voluntary ethanol intake. Using male and female C57BL/6 mice, we explored the causal relationship between hippocampal miR-181a expression and these behaviors. Lentivirus vectors were delivered into the hippocampus for focal miR-181a overexpression in mice. Then behaviors were observed by elevated plus maze (EPM) and open field (OF) tests. Results showed that the viral approach employed to overexpress miR-181a, in the hippocampus, resulted in increased anxiety-like behavior in the EPM and OF tests. Additionally, miR-181a overexpression exacerbated voluntary ethanol intake and preference in the two-bottle choice paradigm without affecting saccharin or quinine consumption. Mechanistically, miR-181a gain-of-function reduced SIRT-1 expression in the hippocampus. These findings demonstrate that miR-181a upregulation in the hippocampus promotes anxiety and ethanol-related behaviors, likely through SIRT-1 repression. This work highlights miR-181a as a key molecular mediator in the epigenetic regulation of mood disorders and ethanol consumption.