Nowadays, cold-formed thin-walled steel tube truss (CTSTT) shear walls are popularly used in engineering practice because of their superiority in light weight, convenient construction and good ductility. However, little attention to the seismic response of the CTSTT shear walls has been paid. This paper aims to investigate the effect of wall openings on seismic behavior of cold-formed thin-walled steel tube truss (CTSTT) shear walls. Five full-scale specimens of CTSTT shear walls sheathed with double-sided oriented strand board (OSB) panels and one full-scale specimen of CTSTT skeleton were tested under vertical axial compression combining lateral low-cyclic load. The parameters, including the type and layout of openings, stressed skin diaphragm as well as opening rate, were estimated in the quasi-static tests. The seismic performance of CTSTT shear wall was evaluated in terms of shear capacity, rigidity, ductility and energy dissipation, etc. Moreover, a simplified empirical formula for predicting the shear capacity of CTSTT shear wall with opening was proposed and verified by the experimental data. The test results indicated that the CTSTT shear wall with double-sided OSB panels exhibited admirable rigidity, good ductility and excellent energy dissipation properties. The shear capacity of CTSTT shear wall decreased by wall opening setting, while seismic behavior of the type of wall could be improved by using four-limb studs within the vicinity of opening. These experimental and analytical studies would improve the application of the CTSTT shear walls in engineering practice.