RNA Therapeutics Institute
Molecular Biology | Nucleic Acids, Nucleotides, and Nucleosides
Reliable detection and quantification of antisense oligonucleotides (ASOs) in experimental and clinical specimens is essential to understand the biological function of novel oligonucleotide-based therapeutics. In this study, we describe a method to detect and quantify ASOs in biological samples, whereby the ASO acts as a splint to direct the ligation of complementary probes and quantitative real-time PCR was used to monitor ligation products. Low levels of 2′-O-MOE gapmer ASO in serum, liver, kidney, lung, heart, muscle, and brain tissues can be detected over a 6-log linear range for detection using this method. This method allows quantification of various types of chemically modified ASOs, including PS linkage, 2′-OMe, 2′-O-MOE, locked nucleic acid (LNA), and siRNA. This method does not require probe modifications, and can be performed using standard laboratory equipment; making it a fast, sensitive, and reliable technique that can be widely applied. This detection method may find potential applications in detection of therapeutic oligonucleotides in biological samples.
Molecular Biology, Antisense oligonucleotides, Quantification, SplintR® ligase, qPCR
Rights and Permissions
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
DOI of Published Version
bioRxiv 2021.06.05.447195; doi: https://doi.org/10.1101/2021.06.05.447195. Link to preprint on bioRxiv.
Shin M, Krishnamurthy PM, Watts JK. (2021). Quantification of Antisense Oligonucleotides by Splint Ligation and Quantitative Polymerase Chain Reaction [preprint]. University of Massachusetts Medical School Faculty Publications. https://doi.org/10.1101/2021.06.05.447195. Retrieved from https://escholarship.umassmed.edu/faculty_pubs/2033
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.