RT Journal Article
SR Electronic
T1 Enhanced Anticoagulation Activity of Functional RNA Origami
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.09.29.319590
DO 10.1101/2020.09.29.319590
A1 Abhichart Krissanaprasit
A1 Carson M. Key
A1 Kristen Froehlich
A1 Sahil Pontula
A1 Emily Mihalko
A1 Daniel M. Dupont
A1 Ebbe S. Andersen
A1 Jørgen Kjems
A1 Ashley C. Brown
A1 Thomas H. LaBean
YR 2020
UL http://biorxiv.org/content/early/2020/10/01/2020.09.29.319590.abstract
AB Anticoagulants are commonly utilized during surgeries and to treat thrombotic diseases like stroke and deep vein thrombosis. However, conventional anticoagulants have serious side-effects, narrow therapeutic windows, and lack safe reversal agents (antidotes). Here, an alternative RNA origami displaying RNA aptamers as target-specific anticoagulant, is described. Improved design and construction techniques for self-folding, single-molecule RNA origami as a platform for displaying pre-selected RNA aptamers with precise orientational and spatial control, are reported. Nuclease resistance is added using 2’-fluoro-modified pyrimidines during in vitro transcription. When four aptamers are displayed on the RNA origami platform, the measured thrombin inhibition and anticoagulation activity is higher than observed for free aptamers, ssRNA-linked RNA aptamers, and RNA origami displaying fewer aptamers. Importantly, thrombin inhibition is immediately switched off by addition of specific reversal agents. Results for ssDNA and ssPNA (peptide nucleic acid) antidotes show restoration of 75% and 95% coagulation activity, respectively. To demonstrate potential for practical, long-term storage for clinical use, RNA origami was freeze-dried, and stored at room temperature. Freshly produced and freeze-dried RNA show identical levels of activity in coagulation assays. Compared to current commercial intravenous anticoagulants, RNA origami-based molecules show promise as safer alternatives with rapid activity switching for future therapeutic applications.Competing Interest StatementThe authors have declared no competing interest.