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The widespread uses of SNAs in diverse chemistry and nanotechnology areas have driven their synthetic innovations toward reduced time\/cost and ever\u2010improved quality. Different from existing methods based on salt\u2010aging, low\u2010pH, freezing, and dehydration conditions, herein a speedy and facile way is reported to produce SNAs in miscible aqueous\u2010alcoholic media with easily adjustable solvent polarities. Critically, it is discovered that the non\u2010aqueous environment of a water\/isopropanol (IPA) mixture plays a vital role in affecting DNA interaction with gold nanoparticles (AuNPs) via different surface\u2010anchoring groups. In addition to significantly accelerated DNA conjugation on AuNPs, the IPA\u2010based strategy achieves various unprecedented controls on the formation of SNAs. These unique features include enhanced DNA adsorption (and ligand desorption) on AuNPs and the possibility of using gold\u2010alkyne linkage for a rapid SNA preparation. Highly efficient SNA syntheses are achieved based on unmodified, thiolated, and alkynated DNA. Moreover, solvent\u2010programmed directionality control of DNA grafts on AuNPs is realized based on alkyne\/phosphorothioate dual\u2010functionalities, which is otherwise unrealizable. The high effectiveness of the IPA\u2010based method points to a previously unconsidered microenvironment\u2010related DNA\u2010nanoparticle interaction for innovating SNA synthesis.<\/jats:p>","DOI":"10.1002\/smll.202507358","type":"journal-article","created":{"date-parts":[[2025,8,25]],"date-time":"2025-08-25T05:10:45Z","timestamp":1756098645000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Advancing Spherical Nucleic Acid Synthesis in Less\u2010Polar Solvents"],"prefix":"10.1002","volume":"21","author":[{"given":"Yichen","family":"Ye","sequence":"first","affiliation":[{"name":"Hefei National Research Center for Physical Sciences at the Microscale Center for Bioanalytical Chemistry School of Chemistry and Materials Science University of Science and Technology of China  Hefei Anhui 230026 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