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Abstract: CO2 flooding is a vital development method for enhanced oil recovery in low-permeability reservoirs. However, micro-fractures are developed in low-permeability reservoirs, which are essential oil flow channels but can also cause severe CO2 gas channeling problems. Therefore, anti-gas channeling is a necessary measure to improve the effect of CO2 flooding. The kind of anti-gas channeling refers to the plugging of fractures in the deep formation to prevent CO2 gas channeling, which is different from the wellbore leakage. Polymer microspheres have the characteristics of controllable deep plugging, which can achieve the profile control of low-permeability fractured reservoirs. In acidic environments with supercritical CO2, traditional polymer microspheres have poor expandability and plugging properties. Based on previous work, a systematic evaluation of the expansion performance, dispersion rheological properties, stability, deep migration, anti-CO2 channeling and enhanced oil recovery ability of a novel acid-resistant polymer microsphere (DCNPM-A) was carried out under CQ oilfield conditions (salinity of 85,000?mg/L, 80?°C, pH?=?3). The results show that the DCNPM-A microsphere had a better expansion performance than the traditional microsphere, with a swelling rate of 13.5. The microsphere dispersion with a concentration of 0.1%–0.5% had the advantages of low viscosity, high dispersion and good injectability in the low permeability fractured core. In the acidic environment of supercritical CO2, DCNPM-A microspheres showed excellent stability and could maintain strength for over 60?d with less loss. In core experiments, DCNPM-A microspheres exhibited delayed swelling characteristics and could effectively plug deep formations. With a plugging rate of 95%, the subsequent enhanced oil recovery of CO2 flooding could reach 21.03%. The experimental results can provide a theoretical basis for anti-CO2 channeling and enhanced oil recovery in low-permeability fractured reservoirs.