3D robotic structures fabricated and powered entirely with proteins

Assembling and upscaling biomolecular activity to perform work in man-made devices constitutes a grand challenge in synthetic biology. Here we report a protocol to construct a fully protein-based micro-3D printed robotic structures that are coated with and actuated by a minimal actomyosin cortex. This protocol describes the detailed procedure including the establishment of 3D printing microstructures with protein materials, the assembly of actomyosin-based active materials, and robotic structure assemblies and quantification. We demonstrated the power of this protocol by programming self-powered soft robots by combining multiple well-investigated biomolecular modules, biophysically analyzing active forces produced in 3D, and engineering smart 3D micro-chips for synthetic cell assembly. Our novel approach offers a versatile platform for constructing large-scale robotics and artificial tissues from bottom-up using individual protein molecules. The detailed step-by-step instructions will guide scientists in replicating the complex preparation procedures, ultimately promoting the advancement and application of 3D protein-based robotic technology.