Scaffold‐based lattice‐engineered 3D DNA origami emerges as a powerful and versatile technique for the rational design and built‐up of arbitrary and monodisperse DNA‐based 3D nanoobjects. Relying on the unsurpassed molecular programmability of sequence‐specific DNA hybridization, a long circular ssDNA strand (termed scaffold) is assembled with many short ssDNA oligomers (termed staples), which organize the scaffold into a 3D lattice in a single step leading to highest precision 3D nanoparticulate structures with high yields. Applications of 3D DNA origami are increasingly wide‐spread and interface with numerous fields of sciences, e.g. anisometric or patchy nanoparticle research, fundamental investigations of superstructure formation, biomedicine, (bio)physics, sensors and optical materials. We will discuss the fundamentals and recent advances from structure formation to selected applications with a mission to promote cross‐disciplinary exchange.