DNA Origami and Its Applications in Synthetic Biology

This review provides a comprehensive summary of recent advancements in DNA origami for synthetic biology, emphasizing two key areas: the precise construction and dynamic regulation of extracellular‐to‐intracellular, and the synergistic integration with cell‐free systems. By combining in vitro assembly with cellular regulation, DNA origami synergizes with cell‐free platforms, opening new pathways for innovative approaches in artificial life design. The review also addresses the challenges and future prospects of this field. Abstract In recent years, DNA origami technology has advanced rapidly as a groundbreaking method for nanomanufacturing. This technology takes advantage of the unique base‐pairing characteristics of DNA, and has significant advantages in constructing spatially ordered and programmable nanostructures. This capability aligns with synthetic biology's core principle of mimicking, extending, and reconstructing natural biological processes by modularly assembling artificial systems. This article provides a comprehensive overview of DNA origami's innovative applications across various domains, including cell membrane surfaces, intercellular communication, intelligent biosensing, and precise gene editing, progressing from the extracellular to the intracellular environment. Finally, this review highlights the synergistic interaction between this technology and cell‐free synthetic biology, achieved through the integration of in vitro assembly and cellular regulation, thereby opening new pathways for the rational design of artificial life systems. This review provides a comprehensive summary of recent advancements in DNA origami for synthetic biology, emphasizing two key areas: the precise construction and dynamic regulation of extracellular-to-intracellular, and the synergistic integration with cell-free systems. By combining in vitro assembly with cellular regulation, DNA origami synergizes with cell-free platforms, opening new pathways for innovative approaches in artificial life design. The review also addresses the challenges and future prospects of this field. Abstract In recent years, DNA origami technology has advanced rapidly as a groundbreaking method for nanomanufacturing. This technology takes advantage of the unique base-pairing characteristics of DNA, and has significant advantages in constructing spatially ordered and programmable nanostructures. This capability aligns with synthetic biology's core principle of mimicking, extending, and reconstructing natural biological processes by modularly assembling artificial systems. This article provides a comprehensive overview of DNA origami's innovative applications across various domains, including cell membrane surfaces, intercellular communication, intelligent biosensing, and precise gene editing, progressing from the extracellular to the intracellular environment. Finally, this review highlights the synergistic interaction between this technology and cell-free synthetic biology, achieved through the integration of in vitro assembly and cellular regulation, thereby opening new pathways for the rational design of artificial life systems. Advanced Science, Volume 13, Issue 2, 9 January 2026.