Recombinant spider silk proteins are characterized by high biocompatibility, biodegradability, anti-inflammatory behaviour and the possibility to adjust the properties to materials applications, making them promising candidates for tissue engineering and biomedical applications. Engineered spider silk proteins inspired by the naturally occurring Araneus diadematus fibroin 4 (ADF4) of the European garden spider, could be produced using a biotechnological process using E. coli ensuring the generation of high, consistent protein amounts. Recombinant eADF4-based spider silk proteins self-assemble into nanofibrils enabling the formation of gels by physical interactions without the need of further crosslinking additives. Due to their good shear-thinning and recovery behaviour gels made of recombinant spider silk proteins are perfectly suited for biofabrication and 3D bioprinting applications to generate constructs exhibiting high shape fidelity. On the one hand these gels could be used as depots for water soluble or insoluble drugs depending on the area of biomedical application. On the other hand spider silk hydrogels could be combined with cells and used for tissue engineering applications and biofabrication of hierarchically structured scaffolds. In this context, the opportunity to genetically modify these spider silk proteins with cell adhesive recognition motifs, such as the integrin-binding peptide RGD, allows a guided material-cell interaction and tissue growth in vitro as well as in vivo. These results show that engineered spider silk materials can be modified and adjusted depending on the application of interest.