{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:58:40Z","timestamp":1785430720971,"version":"3.56.0"},"reference-count":56,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["52303204"],"award-info":[{"award-number":["52303204"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["52303204"],"award-info":[{"award-number":["52303204"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Since the introduction of the spherical nucleic acid (SNA) paradigm in 1996, extensive research has been dedicated to this burgeoning field, yielding groundbreaking advances in biomedicine. Featuring a unique three-dimensional spherical nanoarchitecture composed of highly oriented, densely packed oligonucleotide layers conjugated to a solid or hollow core, SNAs possess extraordinary biological properties, including transfection-reagent-independent cellular internalization and enhanced resistance to nuclease degradation. These synthetic and foundational evolutionary milestones offer profound advantages for the rational design of targeted biomedical therapeutics. In this comprehensive review, recent breakthroughs in the synthetic methodologies and structural designs of SNAs, with a particular emphasis on gold-based templates, are systematically summarized and discussed. Furthermore, the core bottlenecks and future perspectives emerging at the intersection of artificial intelligence and high-throughput screening are highlighted to guide next-generation intelligent nanomedicine development.<\/jats:p>","DOI":"10.3390\/biom16081105","type":"journal-article","created":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T08:16:52Z","timestamp":1785313012000},"page":"1105","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Design of Spherical Nucleic Acids: From Fast Synthesis to Structural Engineering"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2197-4472","authenticated-orcid":false,"given":"Yu","family":"Fan","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering, City University of Hong Kong, Kowloon, Hong Kong SAR 999077, China"},{"name":"Department of Polymeric Materials, School of Materials Science and Engineering, Tongji University, Shanghai 201804, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mei Tsz Jewel","family":"Chan","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, City University of Hong Kong, Kowloon, Hong Kong SAR 999077, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinyuan","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biopharmaceutical Preparation and Delivery, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"12584","DOI":"10.1021\/acs.chemrev.6b00825","article-title":"DNA Origami: Scaffolds for Creating Higher Order Structures","volume":"117","author":"Hong","year":"2017","journal-title":"Chem. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1038\/s41427-023-00470-3","article-title":"Fabricating higher-order functional DNA origami structures to reveal biological processes at multiple scales","volume":"15","author":"Zhou","year":"2023","journal-title":"NPG Asia Mater."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"4416","DOI":"10.1021\/jacs.5b12953","article-title":"Synergy of Two Assembly Languages in DNA Nanostructures: Self-Assembly of Sequence-Defined Polymers on DNA Cages","volume":"138","author":"Chidchob","year":"2016","journal-title":"J. Am. Chem. Soc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1126\/science.1227268","article-title":"Three-Dimensional Structures Self-Assembled from DNA Bricks","volume":"338","author":"Ke","year":"2012","journal-title":"Science"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1038\/382607a0","article-title":"A DNA-based method for rationally assembling nanoparticles into macroscopic materials","volume":"382","author":"Mirkin","year":"1996","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"16291","DOI":"10.1021\/acsnano.3c06564","article-title":"Inspired Beyond Nature: Three Decades of Spherical Nucleic Acids and Colloidal Crystal Engineering with DNA","volume":"17","author":"Mirkin","year":"2023","journal-title":"ACS Nano"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e202400976","DOI":"10.1002\/cbic.202400976","article-title":"Advanced Synthesis of Spherical Nucleic Acids: A Limit-Pursuing Game with Broad Implications","volume":"26","author":"Ye","year":"2025","journal-title":"ChemBioChem"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e120","DOI":"10.1002\/agt2.120","article-title":"Spherical nucleic acids: Organized nucleotide aggregates as versatile nanomedicine","volume":"3","author":"Song","year":"2022","journal-title":"Aggregate"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1021\/acs.accounts.5c00775","article-title":"Spherical Nucleic Acids: Turning Synthetic Advances and Fundamental Discovery into Translational Breakthroughs in Chemistry, Materials Development, Biology, and Medicine","volume":"59","author":"Forsyth","year":"2026","journal-title":"Acc. Chem. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4173","DOI":"10.1002\/smll.201500027","article-title":"The Sequence-Specific Cellular Uptake of Spherical Nucleic Acid Nanoparticle Conjugates","volume":"11","author":"Narayan","year":"2015","journal-title":"Small"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2314232","DOI":"10.1002\/adma.202314232","article-title":"In Vivo Interactions of Nucleic Acid Nanostructures With Cells","volume":"37","author":"Xiao","year":"2025","journal-title":"Adv. Mater."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1376","DOI":"10.1021\/ja209351u","article-title":"Spherical Nucleic Acids","volume":"134","author":"Cutler","year":"2012","journal-title":"J. Am. Chem. Soc."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"8313","DOI":"10.1021\/ac0613582","article-title":"Maximizing DNA Loading on a Range of Gold Nanoparticle Sizes","volume":"78","author":"Hurst","year":"2006","journal-title":"Anal. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"8916","DOI":"10.1021\/ja9719586","article-title":"Characterization of DNA Probes Immobilized on Gold Surfaces","volume":"119","author":"Herne","year":"1997","journal-title":"J. Am. Chem. Soc."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5535","DOI":"10.1021\/ac0006627","article-title":"A Fluorescence-Based Method for Determining the Surface Coverage and Hybridization Efficiency of Thiol-Capped Oligonucleotides Bound to Gold Thin Films and Nanoparticles","volume":"72","author":"Demers","year":"2000","journal-title":"Anal. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1643","DOI":"10.1021\/ja021096v","article-title":"What Controls the Melting Properties of DNA-Linked Gold Nanoparticle Assemblies?","volume":"125","author":"Jin","year":"2003","journal-title":"J. Am. Chem. Soc."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"7266","DOI":"10.1021\/ja3014055","article-title":"Instantaneous and Quantitative Functionalization of Gold Nanoparticles with Thiolated DNA Using a pH-Assisted and Surfactant-Free Route","volume":"134","author":"Zhang","year":"2012","journal-title":"J. Am. Chem. Soc."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6666","DOI":"10.1021\/la0202428","article-title":"Sequence-Dependent Stability of DNA-Modified Gold Nanoparticles","volume":"18","author":"Storhoff","year":"2002","journal-title":"Langmuir"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3896","DOI":"10.1021\/la205036p","article-title":"Surface Science of DNA Adsorption onto Citrate-Capped Gold Nanoparticles","volume":"28","author":"Zhang","year":"2012","journal-title":"Langmuir"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"17053","DOI":"10.1021\/la3035424","article-title":"Instantaneous Attachment of an Ultrahigh Density of Nonthiolated DNA to Gold Nanoparticles and Its Applications","volume":"28","author":"Zhang","year":"2012","journal-title":"Langmuir"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3065","DOI":"10.1021\/jacs.1c00568","article-title":"Flash Synthesis of Spherical Nucleic Acids with Record DNA Density","volume":"143","author":"Hao","year":"2021","journal-title":"J. Am. Chem. Soc."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"9471","DOI":"10.1021\/jacs.7b04885","article-title":"Freezing Directed Construction of Bio\/Nano Interfaces: Reagentless Conjugation, Denser Spherical Nucleic Acids, and Better Nanoflares","volume":"139","author":"Liu","year":"2017","journal-title":"J. Am. Chem. Soc."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2109","DOI":"10.1002\/anie.201814352","article-title":"Freezing-directed Stretching and Alignment of DNA Oligonucleotides","volume":"58","author":"Liu","year":"2019","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2202458","DOI":"10.1002\/smll.202202458","article-title":"Evaporative Drying: A General and Readily Scalable Route to Spherical Nucleic Acids with Quantitative, Fully Tunable, and Record-High DNA Loading","volume":"18","author":"Ye","year":"2022","journal-title":"Small"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e202410353","DOI":"10.1002\/anie.202410353","article-title":"Rapid Thermal Drying Synthesis of Nonthiolated Spherical Nucleic Acids with Stability Rivaling Thiolated DNA","volume":"63","author":"Wang","year":"2024","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1038\/s41467-022-28627-8","article-title":"Fast microwave heating-based one-step synthesis of DNA and RNA modified gold nanoparticles","volume":"13","author":"Huang","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2310026","DOI":"10.1002\/smll.202310026","article-title":"From Structure to Application: The Evolutionary Trajectory of Spherical Nucleic Acids","volume":"20","author":"Wang","year":"2024","journal-title":"Small"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"8459","DOI":"10.1021\/acs.analchem.0c01204","article-title":"A Spherical Nucleic Acid Probe Based on the Au\u2013Se Bond","volume":"92","author":"Gao","year":"2020","journal-title":"Anal. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"5178","DOI":"10.1039\/D0CC01213K","article-title":"Reconstruction of nano-flares based on Au\u2013Se bonds for high-fidelity detection of RNA in living cells","volume":"56","author":"Zhan","year":"2020","journal-title":"Chem. Commun."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4949","DOI":"10.1021\/acs.analchem.6b00891","article-title":"PolyA-Mediated DNA Assembly on Gold Nanoparticles for Thermodynamically Favorable and Rapid Hybridization Analysis","volume":"88","author":"Zhu","year":"2016","journal-title":"Anal. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"781","DOI":"10.1038\/s41563-019-0549-3","article-title":"Programming nanoparticle valence bonds with single-stranded DNA encoders","volume":"19","author":"Yao","year":"2020","journal-title":"Nat. Mater."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"11275","DOI":"10.1021\/acs.analchem.1c02264","article-title":"Precision Spherical Nucleic Acids Enable Sensitive FEN1 Imaging and Controllable Drug Delivery for Cancer-Specific Therapy","volume":"93","author":"Li","year":"2021","journal-title":"Anal. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"9139","DOI":"10.1021\/acs.nanolett.5c02063","article-title":"Circularization of Aptamer Nanomedicine Improves Nuclease Resistance, Targeted Recognition, and Transdermal Effect for Topical Psoriasis Therapy","volume":"25","author":"Liu","year":"2025","journal-title":"Nano Lett."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3867","DOI":"10.1021\/nl3020846","article-title":"Hollow Spherical Nucleic Acids for Intracellular Gene Regulation Based upon Biocompatible Silica Shells","volume":"12","author":"Young","year":"2012","journal-title":"Nano Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"6412","DOI":"10.1021\/acs.nanolett.6c00961","article-title":"Using Well-Defined DNA Nanostructures To Study the Influence of DNA Clustering and Presentation on SNA Cellular Uptake","volume":"26","author":"Ma","year":"2026","journal-title":"Nano Lett."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3174","DOI":"10.1021\/jacs.1c12750","article-title":"Hairpin-like siRNA-Based Spherical Nucleic Acids","volume":"144","author":"Vasher","year":"2022","journal-title":"J. Am. Chem. Soc."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e202416988","DOI":"10.1002\/anie.202416988","article-title":"Tetrahedral DNA Framework-Based Spherical Nucleic Acids for Efficient siRNA Delivery","volume":"64","author":"Li","year":"2025","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e202503004","DOI":"10.1002\/anie.202503004","article-title":"Spherical Nucleic Acids-Directed Cryosynthesis of Manganese Nanoagents for Tumor Imaging and Therapy","volume":"64","author":"Xie","year":"2025","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e2409557122","DOI":"10.1073\/pnas.2409557122","article-title":"cGAS-agonistic spherical nucleic acids reprogram the glioblastoma immune microenvironment and promote antitumor immunity","volume":"122","author":"Mahajan","year":"2025","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4340","DOI":"10.1073\/pnas.1801836115","article-title":"Molecular spherical nucleic acids","volume":"115","author":"Li","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"15843","DOI":"10.1039\/D1SC04833C","article-title":"Molecularly pure miktoarm spherical nucleic acids: Preparation and usage as a scaffold for abiotic intracellular catalysis","volume":"12","author":"Zhang","year":"2021","journal-title":"Chem. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"5043","DOI":"10.1021\/acs.molpharmaceut.3c00370","article-title":"In Vivo Imaging of [60]Fullerene-Based Molecular Spherical Nucleic Acids by Positron Emission Tomography","volume":"20","author":"Auchynnikava","year":"2023","journal-title":"Mol. Pharm."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e202410744","DOI":"10.1002\/anie.202410744","article-title":"Precise Preparation of Supramolecular Spherical Nucleic Acids for Nucleolin-Targeted Gene Delivery","volume":"63","author":"Chen","year":"2024","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1927","DOI":"10.1021\/jacsau.6c00012","article-title":"Host\u2013Guest Assembly of Transmucosal Molecular Spherical Nucleic Acids for Oral Leukoplakia Immunotherapy and Microbiota Homeostasis Restoration","volume":"6","author":"Xu","year":"2026","journal-title":"JACS Au"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"19168","DOI":"10.1002\/anie.202006890","article-title":"Light-Induced Self-Escape of Spherical Nucleic Acid from Endo\/Lysosome for Efficient Non-Cationic Gene Delivery","volume":"59","author":"Shi","year":"2020","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"102562","DOI":"10.1016\/j.xcrp.2025.102562","article-title":"Cyclic brush spherical nucleic acids for enhanced gene-immunotherapy of hepatocellular carcinoma via neutrophil polarization","volume":"6","author":"Liu","year":"2025","journal-title":"Cell Rep. Phys. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3996","DOI":"10.1021\/acsnano.3c05940","article-title":"Control of the Assembly and Disassembly of Spherical Nucleic Acids Is Critical for Enhanced Gene Silencing","volume":"18","author":"Asohan","year":"2024","journal-title":"ACS Nano"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/j.actbio.2024.04.013","article-title":"A fully biodegradable spherical nucleic acid nanoplatform for self-codelivery of doxorubicin and miR122 for innate and adaptive immunity activation","volume":"180","author":"Jiang","year":"2024","journal-title":"Acta Biomater."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1021\/acs.bioconjchem.1c00540","article-title":"An Investigation into the Resistance of Spherical Nucleic Acids against DNA Enzymatic Degradation","volume":"33","author":"Kyriazi","year":"2022","journal-title":"Bioconjugate Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1021\/nl802958f","article-title":"Polyvalent DNA Nanoparticle Conjugates Stabilize Nucleic Acids","volume":"9","author":"Seferos","year":"2009","journal-title":"Nano Lett."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1002\/anie.201409211","article-title":"Spherical Nucleic Acid Nanoparticle Conjugates Enhance G-Quadruplex Formation and Increase Serum Protein Interactions","volume":"54","author":"Chinen","year":"2015","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1603847","DOI":"10.1002\/smll.201603847","article-title":"The Impact of Protein Corona Formation on the Macrophage Cellular Uptake and Biodistribution of Spherical Nucleic Acids","volume":"13","author":"Chinen","year":"2017","journal-title":"Small"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1038\/s41551-019-0351-1","article-title":"Exploration of the nanomedicine-design space with high-throughput screening and machine learning","volume":"3","author":"Yamankurt","year":"2019","journal-title":"Nat. Biomed. Eng."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"15646","DOI":"10.1038\/ncomms15646","article-title":"Real-time visualization of clustering and intracellular transport of gold nanoparticles by correlative imaging","volume":"8","author":"Liu","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1038\/s41596-020-00420-1","article-title":"Tracking endocytosis and intracellular distribution of spherical nucleic acids with correlative single-cell imaging","volume":"16","author":"Liu","year":"2021","journal-title":"Nat. Protoc."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"4517","DOI":"10.1038\/s41467-025-59885-x","article-title":"Nucleic acid spheres for treating capillarisation of liver sinusoidal endothelial cells in liver fibrosis","volume":"16","author":"Liu","year":"2025","journal-title":"Nat. Commun."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/8\/1105\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T08:42:02Z","timestamp":1785314522000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/8\/1105"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,29]]},"references-count":56,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["biom16081105"],"URL":"https:\/\/doi.org\/10.3390\/biom16081105","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,29]]}}}