Biosynthetic DNA origami nanostructures: harnessing E. coli-phage systems as nanofactories

DNA origami nanostructures (DONs) serve as versatile nanocarriers with promising application potential, but high production cost and low yield limit their development. The biosynthetic approach based on the Escherichia coli ( E. coli )-phage system offers an effective route for the low-cost, scalable fabrication of DONs. This approach relies on the biosynthetic production of single-stranded DNA via in vivo rolling circle replication, offering advantages of nucleic acid sequence fidelity and length scalability. This review summarizes recent advances in the biosynthesis of DONs using the E. coli -phage system, with a focus on biosynthetic strategies, and briefly introduces the production of functionalized DONs. Furthermore, we provide a comprehensive overview of the entire biosynthetic workflow, and discuss the advantages and challenges of the biosynthetic approach in depth. This review aims to provide insights for researchers in related fields and to facilitate the development of DNA origami technology.