Lignin, one of the most abundant natural polymers, has been extensively studied as liquid or solid additive in bituminous binders. Despite the fact the organosolv lignin in bitumen improves the overall resistance against oxidative aging, lignin could lead to binders of high thermal cracking sensitivity. In this study, a bio-based oil is implemented in a lignin modified bitumen to ameliorate characteristics, such as fatigue and thermal cracking resistance. Pressure aging vessel conditioning was applied to new binders formulated by different proportions of bio-oil to simulate the long-term aging. A series of rheological tests were performed. Based on the linear amplitude sweep test results, fatigue damage of lignin-bitumen could be reduced by increasing the oil content. According to relaxation test results, the addition of oil significantly decreased the ratio of residual stress and relaxation time. This study has shown preliminary conclusions on the use of bio-oil to improve the low-intermediate temperature performance of lignin-bitumen binders.