DNA Catalyzed Efficient Production of Single-Stranded DNA Nanostructures

Abstract

Single-stranded (ss) DNA nanostructures, created by unimolecular folding of kilo-based ssDNA, represent a novel class of DNA constructs suitable for biomaterial and biomedical applications. Here, using DNA-cleaving DNA enzymes (deoxyribozymes), we present a strategy for efficient and cost-effective production of milligrams of ssDNA nanostructures. We designed “genes” of ssDNA nanostructures flanked by self-cleaving deoxyribozymes and amplified the genes in the single-stranded form. Upon autoprocessing, the resulting amplicons release quadrillion copies of the ssDNA that self-folds into the desired structures. By engineering trans-acting deoxyribozymes, we showed that several ssDNA nanostructures of different shapes can be encoded and amplified in tandem with controllable release of specific shapes from the amplicons, thereby providing a convenient yet economical way to safely store “genetic codes” of nanostructures on an ssDNA for selective mass production on demand. We expect our approach will promote the development of applications of DNA catalysts as well as gene-sized ssDNA in broad areas.