The observation of twinned structures in one-dimensional nanomaterials is very rare. In this paper, we focused our attention on the synthesis of CuS twinned nanorods by a simple wet chemical method, the in situ source−template−interface reaction (ISTIR) route, at low reaction temperature (105 °C). The possible mechanism for the growth of CuS nanorods and nanoparticles was also investigated. The twinned structure of nanorods is discussed in terms of defects introduced during the growth of nanorods. The products have been characterized by X-ray diffraction, energy dispersive X-ray analysis, X-ray photoelectron spectroscopy, and transmission electron spectroscopy analysis. The photoluminescence spectra of twinned nanorods show poor property in comparison to defect-free nanorods.