In this study, a temperature-insensitive strain sensor that detects only the strain without responding to the temperature was designed. The transport mechanism and associated temperature coefficient of resistance (TCR) of a poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) thin film were modified through secondary doping with dimethyl sulfoxide (DMSO). Upon DMSO-doping, the carrier transport mechanism of the PEDOT:PSS thin film transitioned from hopping to band-like transport, with a morphological change. At the DMSO doping level, which caused the critical point of the transport transition, the resistance of the thin film was maintained with a change in temperature. Consequently, the TCR of the optimized PEDOT:PSS thin film was less than 9 × 10-5 K-1, which is 102 times lower than that of the as-prepared films. The carrier mobility of the PEDOT:PSS thin film was effectively improved with the morphological change due to DMSO doping and was investigated through combinational analysis. Ultimately, the wearable strain sensor prepared using the optimized PEDOT:PSS thin film responded stably to the applied strain with a gauge factor of 2 and exhibited excellent temperature anti-interference.