In recent years, significant progress has been made in both three-dimensional (3D) printing technologies and the exploration of silk as an ink to produce biocompatible constructs. Combined with the unlimited design potential of 3D printing, silk can be processed into a broad range of functional materials and devices for various biomedical applications. The ability of silk to be processed into various materials, including solutions, hydrogels, particles, microspheres, and fibers, makes it an excellent candidate for adaptation to different 3D printing techniques. This review presents a didactic overview of the 3D printing of silk-based materials, major categories of printing techniques, and their prototyping mechanisms and structural features. In addition, we provide a roadmap for researchers aiming to incorporate silk printing into their own work by summarizing promising strategies from both technical and material aspects, to relate state-of-the-art silk-based material processing with fast-developing 3D printing technologies. Thus, our focus is on elucidating the techniques and strategies that advance the development of precise assembly strategies for silk-based materials. Precise printing (including high printing resolution, complex structure realization, and printing fidelity) is a prerequisite for the digital design capability of 3D printing technology and would definitely broaden the application era of silk, such as complex biomimetic tissue structures, vasculatures, and transdermal microneedles.