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Volume 14
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10.3390/toxins14070470
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Article

RIPpore: A Novel Host-Derived Method for the Identification of Ricin Intoxication through Oxford Nanopore Direct RNA Sequencing

by
Yan Ryan
1,
Abbie Harrison
1,
Hannah Trivett
1,
Catherine Hartley
1,
Jonathan David
2,
Graeme C. Clark
2,* and
Julian A. Hiscox
1,*
1
Institute for Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L3 5RF, UK
2
Defence Science Technology Laboratory, Salisbury SP4 0JQ, UK
*
Authors to whom correspondence should be addressed.
Toxins 2022, 14(7), 470; https://doi.org/10.3390/toxins14070470
Submission received: 17 June 2022 / Revised: 1 July 2022 / Accepted: 7 July 2022 / Published: 9 July 2022
(This article belongs to the Special Issue Advances in Ricin Antitoxins: From Intoxication to Diagnosis and Treatment)
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Abstract

Ricin is a toxin which enters cells and depurinates an adenine base in the sarcin-ricin loop in the large ribosomal subunit, leading to the inhibition of protein translation and cell death. We postulated that this depurination event could be detected using Oxford Nanopore Technologies (ONT) direct RNA sequencing, detecting a change in charge in the ricin loop. In this study, A549 cells were exposed to ricin for 2–24 h in order to induce depurination. In addition, a novel software tool was developed termed RIPpore that could quantify the adenine modification of ribosomal RNA induced by ricin upon respiratory epithelial cells. We provided demonstrable evidence for the first time that this base change detected is specific to RIP activity using a neutralising antibody against ricin. We believe this represents the first detection of depurination in RNA achieved using ONT sequencers. Collectively, this work highlights the potential for ONT and direct RNA sequencing to detect and quantify depurination events caused by ribosome-inactivating proteins such as ricin. RIPpore could have utility in the evaluation of new treatments and/or in the diagnosis of exposure to ricin.
Keywords: ricin; RNA sequencing; MinIon; ribosome inactivating proteins; saporin; ribosomes ricin; RNA sequencing; MinIon; ribosome inactivating proteins; saporin; ribosomes

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MDPI and ACS Style

Ryan, Y.; Harrison, A.; Trivett, H.; Hartley, C.; David, J.; Clark, G.C.; Hiscox, J.A. RIPpore: A Novel Host-Derived Method for the Identification of Ricin Intoxication through Oxford Nanopore Direct RNA Sequencing. Toxins 2022, 14, 470. https://doi.org/10.3390/toxins14070470

AMA Style

Ryan Y, Harrison A, Trivett H, Hartley C, David J, Clark GC, Hiscox JA. RIPpore: A Novel Host-Derived Method for the Identification of Ricin Intoxication through Oxford Nanopore Direct RNA Sequencing. Toxins. 2022; 14(7):470. https://doi.org/10.3390/toxins14070470

Chicago/Turabian Style

Ryan, Yan, Abbie Harrison, Hannah Trivett, Catherine Hartley, Jonathan David, Graeme C. Clark, and Julian A. Hiscox. 2022. "RIPpore: A Novel Host-Derived Method for the Identification of Ricin Intoxication through Oxford Nanopore Direct RNA Sequencing" Toxins 14, no. 7: 470. https://doi.org/10.3390/toxins14070470

APA Style

Ryan, Y., Harrison, A., Trivett, H., Hartley, C., David, J., Clark, G. C., & Hiscox, J. A. (2022). RIPpore: A Novel Host-Derived Method for the Identification of Ricin Intoxication through Oxford Nanopore Direct RNA Sequencing. Toxins, 14(7), 470. https://doi.org/10.3390/toxins14070470

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