{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T11:53:12Z","timestamp":1781610792634,"version":"3.54.5"},"reference-count":43,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T00:00:00Z","timestamp":1778630400000},"content-version":"vor","delay-in-days":12,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T00:00:00Z","timestamp":1777593600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Amino\u2010functionalized oligonucleotides are valuable tools in therapeutics, biosensing, and nanotechnology. Although 5\u2032\u2010terminal modifications are readily accessible, incorporation at the 3\u2032\u2010terminus remains more challenging and often requires pre\u2010functionalized solid supports or specialized reagents. To address these limitations, we developed a bis(hydroxymethyl)malonyl CPG (CPR II CPG) solid support that can be activated with\n                    <jats:italic>N<\/jats:italic>\n                    ,\n                    <jats:italic>N<\/jats:italic>\n                    \u2032\u2010disuccinimidyl carbonate (DSC) to generate a carbamate\u2010forming platform. Using this support, we established an on\u2010support bis\u2010carbamate construction strategy that enables efficient incorporation of amino modifiers in high yields and crude purities. Furthermore, by leveraging a trifunctional linker (1\u2010amino\u20103\u2010(((4\u2010methoxyphenyl)diphenylmethyl)amino)propan\u20102\u2010ol), we developed a modular approach in which one amine is tethered to the solid support while the second amine can be selectively detritylated and conjugated to diverse ligands. The free hydroxyl group can then be used to extend an oligonucleotide through automated solid\u2010phase synthesis. This strategy accommodates a broad range of ligands through multiple conjugation chemistries, including NHS esters\/carbonates and EDC\u2010mediated amide bond formation, with good conversions. Finally, we describe a tandem oligonucleotide synthesis (TOS) protocol for generating strands of DNA bearing ligands at defined terminal positions, enabling rapid access to highly modified oligonucleotides. All oligonucleotide conjugates were purified using standard workflows, including strong anion exchange high\u2010performance liquid chromatography (SAX\u2010HPLC), and characterized by mass spectrometry (MS). \u00a9 2026 The Author(s).\n                    <jats:italic>Current Protocols<\/jats:italic>\n                    published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Synthesis of\n                    <jats:bold>\n                      <jats:italic>R<\/jats:italic>\n                    <\/jats:bold>\n                    \u2010CPG and incorporation of bis\u2010amino modifiers such as putrescine\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Synthesis of MMTr\u20101,3\u2010diamino\u20102\u2010propanol (compound\n                    <jats:bold>2<\/jats:bold>\n                    ) and\n                    <jats:bold>\n                      <jats:italic>R<\/jats:italic>\n                    <\/jats:bold>\n                    \u2010CPG functionalization\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol 1<\/jats:bold>\n                    : EDC\u2010mediated ligand\u2010amine coupling (4\u2010bromobenzoic acid) and DNA synthesis\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol 2<\/jats:bold>\n                    : NHS\u2010carbonate\u2010mediated ligand coupling (Boc\u2010OSu) and RNA synthesis\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : Conjugational tandem oligonucleotide synthesis (cTOS) of highly modified DNA duplexes\n                  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