Gas turbine blades normally operate under high-temperatures, extreme pressure, and high-speed conditions, which leads to complex failure mechanisms and difficulties in fatigue life assessment. Moreover, due to the randomness of manufacturing errors, working loads, and material properties, the low cycle fatigue (LCF) life normally presents unavoidable stochastic behavior. Therefore, accurate and efficient fatigue life prediction is critical for the design of gas turbine blades. Accordingly, this paper analyzes the effects of mean stress, anisotropy, and uncertainties on the LCF life of nickel-based single-crystal turbine blades. The modified Hill yield criterion is employed to deal with the anisotropic damage characteristics of nickel-based single-crystal superalloy (NSCS) under a multiaxial stress state. Meanwhile, the effects of the temperature and crystal orientation are introduced into the walker mean stress correction term. A multiaxial LCF life prediction model is developed based on the energy criterion-based fatigue life estimation method. Moreover, multi-source uncertainties originating from rotation speeds, material properties, and temperature are analyzed and quantified. Comparison between the predicted and tested life indicates that the proposed method produces good accuracy and robustness, which can provide a reference for the reliability design of NSCS turbine blades.