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Sitobion miscanthiis a wheat aphid species that can damage seriously agricultural production. The effective management of wheat aphids has depended on chemical insecticides. However, their wide application led to severe resistance of wheat aphids to some insecticides, and cytochrome P450 as a detoxifying enzyme plays a crucial role in the insecticide resistance. In this study, CYP6CY1, a new P450 gene, was isolated and overexpressed in a S.miscanthiresistant strain to imidacloprid. The increased sensitivity to imidacloprid after silencing of CYP6CY1indicated that its involvement in imidacloprid resistance. Subsequently, action of miR-3047-3p within the 3′ UTR of the CYP6CY1was confirmed at the posttranscriptional regulatory level, and was shown to be involved in the resistance to imidacloprid. These results provide opportunities for understanding the roles of P450 in insecticide resistance of pests.