Natural silk fiber derived from the Bombyx mori (B. mori) silkworm has long been used as a luxury raw material in textile industry because of its shimmering appearance and durability, and as surgical suture for its high strength and flexibility. Regenerated silk fibroin, as the main protein extracted from the cocoons of the B. mori silkworm, recently has gained considerable attention due to its outstanding properties, including facile processability, superior biocompatibility, controllable biodegradation, and versatile functionalization. Tremendous effort has been made to fabricate silk fibroin into various promising materials with controlled structural and functional characteristics for advanced utilities in a multitude of biomedical applications, flexible optics, electronics devices, and filtration systems. Herein, reverse engineered silk fibroin extraction methods are reviewed, recent advances in extraction techniques are discussed. Fabrication methods of silk fibroin materials in various formats are also addressed in detail; in particular, progress in new fabrication technologies is presented. Attractive applications of silk fibroin-based materials are then summarized and highlighted. The challenges faced by current approaches in production of silk fibroin-based materials and future directions acquired for pushing these favorable materials further toward above mentioned applications are further elaborated.