Saraya Jagandeep S, Horton Nicholas G, Capperauld Michael J, Zakaria Evan, O'Flaherty Derek K
Department of Chemistry, University of Guelph, 50 Stone Rd E, Guelph, Ontario, N1G 2W1, Canada.
Chem Asian J. 2025 Aug;20(16):e00537. doi: 10.1002/asia.202500537. Epub 2025 Jun 2.
Chemical modifications of oligonucleotides are routinely employed to enhance their functional properties. Amino-modifiers serve as versatile chemical handles for postsynthetic (bio)conjugation, nucleic acid immobilization on solid supports, and investigations into nonenzymatic genome replication relevant to the origins of life, to name a few. Here, we report a cost-effective, disulfide-containing solid-support linkage that enables the on-column synthesis of nucleic acids with 3'-amino or 3'-phosphate modifications. The orthogonality of this solid-support linker facilitates an on-column protecting group strategy, enabling the synthesis of DNA and RNA containing 3'-amino-2',3'-dideoxyribosides from commercial unprotected mononucleosides. Additionally, we present an on-column deprotection protocol for DNA and RNA, prior to cleavage from the solid support, eliminating the precipitation step typically required in conventional RNA workflows, leading to higher recovery for certain strands. Expanding on our previous work, we introduce a versatile modular tandem oligonucleotide synthesis (mTOS) approach, allowing selective release of downstream strands from the one directly bound to the solid-support via the disulfide-containing linker. Together, these advances in solid-support design and oligonucleotide synthesis unlock new opportunities in bioconjugation, biotechnology, and the study of prebiotic replication mechanisms, broadening the utility of chemically modified nucleic acids across research disciplines.
寡核苷酸的化学修饰经常被用于增强其功能特性。氨基修饰剂可作为通用的化学手柄，用于合成后（生物）共轭、核酸在固体支持物上的固定以及与生命起源相关的非酶促基因组复制研究等。在此，我们报告一种经济高效的含二硫键的固体支持物连接方式，它能够实现对具有3'-氨基或3'-磷酸修饰的核酸进行柱上合成。这种固体支持物连接子的正交性有助于实施柱上保护基团策略，从而能够从市售的未保护单核苷合成含有3'-氨基-2',3'-二脱氧核糖核苷的DNA和RNA。此外，我们还提出了一种在从固体支持物上切割之前对DNA和RNA进行柱上脱保护的方案，省去了传统RNA工作流程中通常需要的沉淀步骤，从而提高了某些链的回收率。在我们之前工作的基础上进行拓展，我们引入了一种通用的模块化串联寡核苷酸合成（mTOS）方法，允许通过含二硫键的连接子从直接与固体支持物结合的链上选择性释放下游链。总之，这些在固体支持物设计和寡核苷酸合成方面的进展为生物共轭、生物技术以及益生元复制机制研究带来了新机遇，拓宽了化学修饰核酸在各研究领域的应用范围。
