University of Southern Denmark, Department of Physics, Chemistry and Pharmacy, Campusvej 55, 5230 Odense M, Denmark.
Org Biomol Chem. 2016 Aug 7;14(29):6985-95. doi: 10.1039/c6ob01120a. Epub 2016 Jun 29.
Assays for mismatch discrimination and detection of single nucleotide variations by hybridization-controlled assembly of liposomes, which do not require tedious surface chemistry, are versatile for both DNA and RNA targets. We report herein a comprehensive study on different DNA and LNA (locked nucleic acids) probe designs, including membrane-anchoring requirements, studies on different probes and target lengths (including overhangs), DNA and RNA targets (including sequences associated with pathogens) for lipidated nucleic acids (LiNAs). Advantages and limitations of the liposome assembly based assay in the context of mismatch discrimination and SNP detection are presented. The advantages of membrane-anchored LiNA-probes compared to chemically attached probes on solid nanoparticles (e.g. gold nanoparticles) are described. Key functionalities such as non-covalent attachment of LiNA probes without the need for long spacers and the inherent mobility of membrane-anchored probes in lipid-bilayer membranes will be described for several different probe designs.
通过脂质体的杂交控制组装进行错配识别和单核苷酸变异检测的测定法,不需要繁琐的表面化学处理,对 DNA 和 RNA 靶标都具有多功能性。我们在此报告了对不同 DNA 和 LNA(锁核酸)探针设计的全面研究,包括膜锚定要求、不同探针和靶标长度(包括突出端)、脂质化核酸(LiNAs)的 DNA 和 RNA 靶标(包括与病原体相关的序列)的研究。介绍了基于脂质体组装的测定法在错配识别和 SNP 检测方面的优缺点。与化学附着在固体纳米颗粒(例如金纳米颗粒)上的探针相比,描述了膜锚定 LiNA 探针的优势。将描述几种不同探针设计的关键功能,例如无需长间隔物的 LiNA 探针的非共价附着以及膜锚定探针在脂质双层膜中的固有迁移性。