The development of real-time trace-level NO2 quantification platforms that can be operated at room temperature constitutes a critical advancement for occupational safety and public health monitoring systems. This study demonstrates a room-temperature NO2 sensor using MXene-Cu2O composites prepared via a hydrothermal method. Systematic evaluation of MXene-introduced effects identified the 0.84 wt % MXene-Cu2O composite as optimal, exhibiting 4-fold enhanced sensitivity and shorter response (55 s)/recovery (35 s) time compared to pure Cu2O. Additionally, the sensor exhibits a low detection limit (10 ppb), high selectivity, great reversibility, and long-term stability. The enhanced sensing performance originates from precisely engineered interfacial architectures between MXene and Cu2O, which effectively adjust the charge-transfer behavior through the conduction tunnel in the sensing material. Furthermore, oxygen vacancy engineering creates defect-mediated adsorption centers that promote selective NO2 chemisorption through charge polarization effects. This research offers a novel strategy for designing optimized structures to enhance the sensitivity of MOS-based materials for NO2 gas detection.