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dc.contributor.authorShin, Minwook
dc.contributor.authorKrishnamurthy, Pranathi Meda
dc.contributor.authorWatts, Jonathan K
dc.date2022-08-11T08:08:27.000
dc.date.accessioned2022-08-23T15:55:46Z
dc.date.available2022-08-23T15:55:46Z
dc.date.issued2021-06-05
dc.date.submitted2021-07-14
dc.identifier.citation<p>bioRxiv 2021.06.05.447195; doi: https://doi.org/10.1101/2021.06.05.447195. <a href="https://doi.org/10.1101/2021.06.05.447195" target="_blank" title="view preprint in biorxiv">Link to preprint on bioRxiv.</a></p>
dc.identifier.doi10.1101/2021.06.05.447195
dc.identifier.urihttp://hdl.handle.net/20.500.14038/29826
dc.description<p>This article is a preprint. Preprints are preliminary reports of work that have not been certified by peer review.</p>
dc.description.abstractReliable 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.
dc.language.isoen_US
dc.relationNow published in Nucleic Acid Therapeutics doi: 10.1089/nat.2021.0040
dc.rightsThe 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.
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectMolecular Biology
dc.subjectAntisense oligonucleotides
dc.subjectQuantification
dc.subjectSplintR® ligase
dc.subjectqPCR
dc.subjectMolecular Biology
dc.subjectNucleic Acids, Nucleotides, and Nucleosides
dc.titleQuantification of Antisense Oligonucleotides by Splint Ligation and Quantitative Polymerase Chain Reaction [preprint]
dc.typePreprint
dc.source.journaltitlebioRxiv
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=3061&amp;context=faculty_pubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/faculty_pubs/2033
dc.identifier.contextkey23822836
refterms.dateFOA2022-08-23T15:55:46Z
html.description.abstract<p><p id="x-x-x-p-2">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′-<em>O</em>-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′-<em>O</em>-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.</p>
dc.identifier.submissionpathfaculty_pubs/2033
dc.contributor.departmentRNA Therapeutics Institute


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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.
Except where otherwise noted, this item's license is described as 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.