Wilbanks Brandon A, Doherty Caroline D, Pearson Keenan S, Jain Sonia, Sarkaria Jann N, Maher L James
Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine and Science, Rochester, MN 55905, USA.
Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic Graduate School of Biomedical Sciences, Rochester, MN 55905, USA.
Mol Ther Methods Clin Dev. 2025 Apr 28;33(2):101482. doi: 10.1016/j.omtm.2025.101482. eCollection 2025 Jun 12.
Aptamers are folded single-stranded DNA or RNA molecules with high affinity for target small molecules, proteins, cells, or tissues. Affinity arises from the three-dimensional structure of the aptamer. Many significant advancements in oligonucleotide selection strategies have been made since the first foundational reports of selection techniques over 30 years ago. Most notably, the last 15 years have seen major advancements in selection protocols for aptamers with affinity to unspecified targets in complex living mammalian contexts such as cultured cells or tissues and tumors in live animals. Chemically unmodified DNA aptamers have potential as low-cost, biodegradable drug delivery or diagnostic reagents offering highly specific interactions against targeted cell populations. Here, we provide routine protocols from our labs for design and implementation of aptamer selections against cells or tissues, including principles of library design, selection strategy, aptamer candidate identification, and complementary image- and qPCR-based techniques for validating aptamer specificity.
适体是对靶小分子、蛋白质、细胞或组织具有高亲和力的折叠单链DNA或RNA分子。亲和力源于适体的三维结构。自30多年前首次发表关于筛选技术的基础报告以来，寡核苷酸选择策略已取得许多重大进展。最值得注意的是，在过去15年中，针对复杂活体哺乳动物环境（如培养细胞、组织和活体动物肿瘤）中未指定靶标的适体选择方案取得了重大进展。化学未修饰的DNA适体有潜力作为低成本、可生物降解的药物递送或诊断试剂，针对靶向细胞群体提供高度特异性的相互作用。在这里，我们提供了我们实验室针对细胞或组织进行适体选择的设计和实施的常规方案，包括文库设计原则、选择策略、适体候选物鉴定，以及用于验证适体特异性的基于图像和qPCR的互补技术。
