Programmable DNA-protein building blocks were created by attaching DNA strands to traptavidin, enabling the directed self-assembly of supramolecular structures, such as chains, clusters, and lattices, through sequence-specific DNA hybridization.

Intracellular delivery of folded, functional proteins remains a critical barrier to realizing the full potential of protein therapeutics. We present a fully aqueous, genetically encoded platform for assembling micellar protein nanoparticles via electrostatic coacervation between anionic globular proteins and multi-domain intrinsically disordered proteins (IDPs) composed of a neutral elastin-like polypeptide (ELP) domain and a cationic disordered histone-derived domain (H5). This biosynthetic system allows modular control over charge density, neutral domain length, and stoichiometry, enabling the formation of homogeneous micellar nanoparticles with high protein payload retention under mild, biologically relevant conditions without organic solvents or covalent modification. Unlike amphiphilic micelle systems, globular proteins in this approach act as both cargo and assembly drivers, providing a direct handle to tune encapsulation. Nanoparticle assembly was characterized with a suite of complementary techniques including DLS, FCS, TEM, and SAXS, enabling detailed analysis of nanoparticle structure, size, and composition. The system exhibits remarkable tolerance to different formulations, consistently producing well-defined particles with low dispersity. This expanded design space arises from the use of structured, charge-regulating proteins and confers exceptional versatility in formulation. Different charge fractions modulate not only nanoparticle size but also physicochemical properties such as core density and cargo loading, which may be critical for tuning nanoparticle stability and performance. Together, these features establish a robust and programmable platform for protein-based nanoparticle engineering with potential application in intracellular protein delivery.

Nanoscale analogs of atomic valence enable the rationalization of nanoparticle self-assembly through defined directional interactions. For nanoparticles and colloids, the patchy-particle representation captures the essence of directional interactions; however, it significantly simplifies molecular details and can fail to depict molecular-level contributions to binding modes and consequent assembly behavior. Here, using a system of self-assembled DNA-origami wireframes, we reveal a molecular mechanism that unexpectedly controls inter-patch interactions leading to either monogamous (one-to-one) or polygamous (multi-patch) binding modes, resulting in different superlattice structures with periodic phase behavior. The intrinsic helicity of the double-stranded (ds) DNA linker that connects the wireframes sets the rotational register between patches. The magnitude of helical rotation depends on the length of dsDNA and is sensitive even to a single nucleotide. We discovered that small changes in linker length drastically alter inter-patch orientations, inducing either monogamous or polygamous interactions with different bond topologies and periodic transformation of self-assembled phases. Using X-ray scattering and electron microscopy, we observed periodic switching among simple cubic, body-centered tetragonal, and face-centered cubic lattices for octahedral wireframes, and between body-centered cubic and face-centered cubic lattices for cubic wireframes. Molecular dynamics simulations of the self-assembly process reveal that the observed phase behavior is governed by the relative patch orientations, which are experimentally controlled by the linkers. Our results provide molecular insights into the emergence of topologically complex binding modes and offer levers to control the organization of nanomaterials through synchronization with molecular helicity.