Dynamic traffic flow, which can facilitate the efficient operation of traffic road networks, is an important prerequisite for the application of reasonable assignment of traffic demands in an urban road network. In order to improve the accuracy of dynamic traffic flow assignment, this paper proposes a dynamic traffic flow assignment model based on GPS trajectory data and the influence of POI. First, this paper explores the impact patterns of POI on regional road network congestion during peak hours through qualitative and quantitative analysis. Then, based on the user equilibrium theory, a dynamic traffic flow assignment model, in which the effect of POI on links is reflected using the link-node impedance function, is proposed. Finally, the accuracy of the model is validated by the GPS trajectory data and origin–destination (OD) traffic data of motor vehicles in Xuhui District, Shanghai, China. The results show that the model can be used to coordinate and optimize the traffic assignment of the regional road network under the influence of POI during peak hours and alleviate the congestion of the road network. The findings can provide a powerful reference for developing scientific and rational traffic assignment decisions and management strategies for urban road network traffic.