Structural coloration of photonic crystals (PCs) is considered an ecological and environmental way to achieve colorful textiles. However, constructing PCs with obvious structural colors on traditional flexible yarns is still a great challenge. As a secondary structure that forms textiles, compared with fibers and fabrics, the yarns are rougher, hindering the construction of regular PCs. In this work, the flexible acrylic yarns with vivid structural colors, named PC-based structural color yarns, were prepared by constructing regular PCs via assembling poly(styrene-butyl acrylate-methacrylate) (P(St-BA-MAA)) colloidal microspheres on yarns. Specifically, the properties of P(St-BA-MAA) colloidal microspheres were investigated. The PCs with better structural stability and obvious structural colors were prepared by presetting the acrylic adhesive layer on yarns. Moreover, the color durability and color regulation methods of prepared PC-based structural color yarns were evaluated and discussed. The results showed that the P(St-BA-MAA) colloidal microspheres exhibited even particle sizes, excellent monodispersity, and a typical hard core−soft shell structure. And the glass-transition temperature (T g ) of the microspheres was tested to be about 65.6 °C. The cationic acrylate regarded as a pretreatment agent could not only improve the combination between the PC layers and the yarns by acting as a "bridge" but also enhance the structural color effect by smoothing the yarn surface. The results showed that when the mass fraction of cationic acrylate was 3 wt %, the microspheres were beneficial to access regular PCs with obvious structural colors. The PCs with bright structural colors could be constructed on black acrylic yarns, and the colors of yarns were still bright after rubbing and washing tests, indicating that the prepared PC-based structural color yarns have good color fastness. Moreover, the color hue of PC-based structural color yarns could be regulated by adjusting the particle sizes and viewing angles. This study provides strategic support for the structural coloration of flexible materials.