Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zürich, Zürich, Switzerland.
Department of Paediatrics, Medical Sciences Division, University of Oxford, Oxford, UK.
Methods Mol Biol. 2022;2537:335-350. doi: 10.1007/978-1-0716-2521-7_19.
We describe methods to follow the fate of oligonucleotides after their injection into experimental animals. The quantitation in various tissues, blood or bone marrow cells is possible by chemical ligation PCR. This method works independently of chemical modifications of the oligonucleotide and/or its conjugations to lipid or peptide moieties. Moreover, metabolization intermediates can be detected by mass spectrometry. Together with a readout assay for the biochemical or physiological effects, which will differ, depending on the particular purpose of the oligonucleotide, these methods allow for a comprehensive understanding of oligonucleotide behavior in a living organism.
我们描述了在将寡核苷酸注射到实验动物体内后跟踪其命运的方法。通过化学连接 PCR 可以定量检测各种组织、血液或骨髓细胞中的寡核苷酸。该方法独立于寡核苷酸的化学修饰及其与脂质或肽部分的缀合。此外，还可以通过质谱检测代谢产物中间体。结合用于生化或生理效应的读出测定法（其将根据寡核苷酸的特定用途而有所不同），这些方法可使我们全面了解寡核苷酸在生物体中的行为。
