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Chemosensory proteins (CSPs) are a class of transporters in arthropods. Deeper research on CSPs showed that CSPs may be involved in some physiological processes beyond chemoreception, such as insect resistance to pesticides. We identified two upregulated <i>CSPs</i> in two resistant strains of <i>Aphis gossypii</i> Glover. To understand their role in the resistance of aphids to pesticides, we performed the functional verification of <i>CSP1</i> and <i>CSP4</i> in vivo and in vitro. Results showed that the sensitivity of the thiamethoxam-resistant strain to thiamethoxam increased significantly with the silencing of <i>CSP1</i> and <i>CSP4</i> by RNAi (RNA interference), and the sensitivity of the spirotetramat-resistant strain to spirotetramat increased significantly with the silencing of <i>CSP4</i>. Transgenic <i>Drosophila melanogaster</i> expressing <i>CSPs</i> exhibited stronger resistance to thiamethoxam, spirotetramat, and alpha-cypermethrin than the control did. In the bioassay of transgenic <i>Drosophila</i>, <i>CSPs</i> showed different tolerance mechanisms for different pesticides, and the overexpressed <i>CSPs</i> may play a role in processes other than resistance to pesticides. In brief, the present results prove that <i>CSPs</i> are related to the resistance of cotton aphids to insecticides.