Bijan Zakeri
02-­�06-­�15

SpyTag/SpyCatcher: programmable protein assembly for synthetic biology

Bijan Zakeri, Allen Y. Chen, Zhengtao Deng, Peter A. Carr, and Timothy K. Lu



Protein-­�protein interactions in biology are generally weak and temporary.
Yet many applications in synthetic biology require the stable and long-­�term assembly of proteins post-­�translationally. Conventional assembly methods that rely on non-­�covalent interactions lead to the engineering of unstable and transient molecular architectures that are not ideal for most downstream applications. Here we present the application of the SpyTag/SpyCatcher covalent tagging system for genetically programmable and irreversible engineering of covalent protein architectures both in vitro and in vivo. We demonstrate covalent protein assembly for engineering living materials using biofilms and nanoparticles that bridge the biotic-­�abiotic interphase. Furthermore, we demonstrate the intracellular assembly of a resistance marker for the maintenance of dual plasmids with a single origin, for applications in molecular steganography for DNA camouflage.