Changes in microRNA (miRNA)-mediated gene expression in the nucleus accumbens (NAc) may play important roles in regulating drug addiction. MiR-29c is a highly expressed miRNA in the human and rodent nervous systems where it plays a broad regulatory role. As the first step towards investigating potential functions of miR-29c in methamphetamine (METH) addiction, we used C57BL/6 mice in a model of METH-induced locomotor sensitization. We measured miR-29c expression changes in the NAc of the mice after repeated-intermittent METH exposure and acute METH administration respectively by using quantitative real-time PCR (qPCR). We found that miR-29c expression was significantly down-regulated in the NAc of METH-sensitized mice but not in the acute METH-treated mice. Then, we tested the respective effects of miR-29c over-expression and inhibition in the NAc on METH-induced locomotor sensitization. To reach this goal, we constructed adeno-associated virus (AAV)-expressing miR-29c (AAV-miR-29c) and its corresponding inhibitor - tough decoy (AAV-anti-miR-29c TuD) to over-express and inhibit miR-29c, respectively. We found that AAV-miR-29c over-expression in the NAc enhanced METH-induced locomotor sensitization, whereas AAV inhibition of miR-29c expression in the NAc attenuated the effects of METH. Moreover, we observed the participation of Dnmt3a, Dnmt3b, and Meg3 in the effects of miR-29c on METH sensitization. Our results suggest that miR-29c is an important epigenetic regulator of METH-induced behavioural sensitization and changes in gene expression. These data further suggest a potential role of miR-29c in regulating long-term METH-induced adaptation in the brain.