Silk fibroin (SF) is a structural protein derived from natural silkworm silks. Materials fabricated based on SF usually inherit extraordinary physical and biological properties, including high mechanical strength, toughness, optical transparency, tailorable biodegradability, and biocompatibility. Therefore, SF has attracted interest in the development of sustainable biodevices, especially for emergent bio-electronic technologies. To expand the function of current silk devices, the SF characteristic sequence has been used to synthesize recombinant silk proteins that benefit from SF and other functional peptides, such as stimuli-responsive elastin peptides. In addition to genetic engineering methods, innovated chemistry modification approaches and improved material processing techniques have also been developed for fabricating advanced silk materials with tailored chemical features and nanostructures. Herein, this review summarizes various methods to synthesize functional silk-based materials from different perspectives. This review also highlights the recent advances in the applications of natural and recombinant silks in tissue regeneration, soft robotics, and biosensors, using B. mori SF and silk-elastin-like proteins (SELPs) as examples.