Negative Poisson’s ratio (NPR) material with unique geometry is rare in nature and has an auxetic response under strain in a specific direction. With this unique property, this type of material is significantly promising in many specific application fields. The curling structure commonly exists in knitted products due to the unbalanced force inside a knit loop. Thus, knitted fabric is an ideal candidate to mimic natural NPR materials, since it possesses such an inherent curly configuration and the flexibility to design and process. In this work, a weft-knitted Miura-ori fold (WMF) fabric was produced that creates a self-folding three-dimensional structure with NPR performance. Also, a finite element analysis model was developed to simulate the structural auxetic response to understand the deformation mechanism of hierarchical thread-based auxetic fabrics. The simulated strain–force curves of four WMF fabrics quantitatively agree with our experimental results. The auxetic morphologies, Poisson’s ratio and damping capacity were discussed, revealing the deformation mechanism of the WMF fabrics. This study thus provides a fundamental framework for mechanical-stimulating textiles. The developed NPR knitted fabrics have a high potential to be employed in areas of tissue engineering, such as artificial blood vessels and artificial folding mucosa.