Nature has evolved a wide range of strategies to create self-assembled protein nanostructures with structurally defined architectures that serve a myriad of highly specialized biological functions. With the advent of biological tools for site-specific protein modifications and de novo protein design, a wide range of customized protein nanocarriers have been created using both natural and synthetic biological building blocks to mimic these native designs for targeted biomedical applications. In this review, different design frameworks and synthetic decoration strategies for achieving these functional protein nanostructures are summarized. Key attributes of these designer protein nanostructures, their unique functions, and their impact on biosensing and therapeutic applications are discussed.

We report a new approach for conditional cellulosome formation using toehold-gated dCas9-guided protein assembly. Binding of SpdCas9-CBD is activated by a RNA trigger to unblock the spacer region of thgRNA by toehold-mediated strand displacement.

We report a new modular strategy to assemble dCas9-guided enzyme cascades by employing orthogonal post-translation chemistry. Two orthogonal SpyCatcher and SnoopCatcher pairs were used for the one-pot enzyme bioconjugation onto...

The wide variety of enzymatic pathways that can benefit from enzyme scaffolding is astronomical. While enzyme co‐localization based on protein, DNA, and RNA scaffolds has been reported, we still lack scaffolds that offer well‐defined and uniform three‐dimensional structures for enzyme organization. Here we reported a new approach for protein co‐localization using naturally occurring protein nanocages as a scaffold. Two different nanocages, the 25 nm E2 and the 34 nm heptatitis B virus, were used to demonstrate the successfully co‐localization of the endoglucanase CelA and cellulose binding domain using the robust SpyTag/SpyCatcher bioconjugation chemistry. Because of the simplicity of the assembly, this strategy is useful not only for in vivo enzyme cascading but also the potential for in vivo applications as well.