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.
Keywords: Chemical ligation qPCR; Mass spectrometry; Oligonucleotide biodistribution; Oligonucleotide conjugates; Oligonucleotide therapeutics.
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.