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Using the reliable Watson\u2013Crick base-pairing, scientists have created DNA structures that are compatible with living systems, precisely arranged, and responsive to changes. This article reviews a comprehensive examination over DNA origami, beginning with its conceptual origin and structural evolution, followed by a comparative classification of key nanostructures such as tiles, scaffolded origami, DNA bricks, and single-stranded origami. Advanced design methodologies supported by algorithmic tools, including caDNAno, MagicDNA, DAEDALUS, and oxDNA simulations, are also analyzed. Functionalization strategies are categorized into pre- and post-assembly approaches, highlighting chemical conjugation techniques for targeting and therapeutic loading. A significant focus is on stimuli-responsive systems that react with biological or physical factors such as pH, enzymes, redox microenvironment, and light. Clinical applications are explored through case studies in drug delivery, biosensing, immunotherapy, and gene regulation. Furthermore, challenges, including\n                    <jats:italic>in vivo<\/jats:italic>\n                    stability, immune response, and regulatory uncertainty have been critically assessed; while proposing future directions such as hybrid materials and personalized nanomedicine. Ultimately, a comprehensive overview of DNA origami mechanisms evolve into clinically relevant nanoplatforms for precision therapy and diagnostic tools is provided.\n                  <\/jats:p>","DOI":"10.1088\/1361-6528\/aeae45","type":"journal-article","created":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T22:48:57Z","timestamp":1790808537000},"page":"432002","update-policy":"https:\/\/doi.org\/10.1088\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["DNA nanostructures: design, functionalization, applications, and AI-driven advances"],"prefix":"10.1088","volume":"37","author":[{"given":"Seyed Mohammad Sina","family":"Alemohammad","sequence":"first","affiliation":[{"name":"Ahvaz Jundishapur University of Medical Sciences"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Marziyeh","family":"Mousazadeh","sequence":"additional","affiliation":[{"name":"Tarbiat Modares University"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Zahra","family":"Saremi","sequence":"additional","affiliation":[{"name":"Lorestan University of Medical Sciences"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Mohammad","family":"Rahmanian","sequence":"additional","affiliation":[{"name":"Shahid Beheshti University of Medical Sciences"},{"name":"Shahid Beheshti University of Medical Sciences"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6742-1323","authenticated-orcid":true,"given":"Hossein","family":"Soltaninejad","sequence":"additional","affiliation":[{"name":"Tarbiat Modares University"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]}],"member":"266","published-online":{"date-parts":[[2026,10,23]]},"reference":[{"key":"nanoaeae45bib1","doi-asserted-by":"publisher","first-page":"7882","DOI":"10.1007\/s12010-025-05415-w","type":"journal-article","article-title":"A chimeric peptide-carbon quantum dot nanobiohybrid system for gene delivery to macrophage cells","volume":"197","author":"Mousazadeh","year":"2025","journal-title":"Appl. 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