Golden Gate Assembly of DNA Nanostructures

Golden Gate assembly is a popular molecular cloning method to assemble multiple DNA fragments in a “one-pot” enzymatic reaction. Taking advantage of the high combination efficiency provided by the enzyme cocktail of this assembly method, we present a controllable assembly system of synthetic DNA nanostructures in this work. Similar to the rendering of duplex fragments to be assembled in Golden Gate assembly, type II restriction enzyme creates specific sticky end pairs for the DNA nanostructure units to connect, and ligase consolidates the sticky end cohesion, which collectively enables the corresponding hierarchical assembly. By application of this molecular cloning method in DNA self-assembly, the enzymatic treatment leads to the production of a number of high-order DNA nanostructures.