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

Ryan, Yan ORCID: 0000-0002-4708-5350, Harrison, Abbie, Trivett, Hannah, Hartley, Catherine, David, Jonathan, Clark, Graeme C and Hiscox, Julian A ORCID: 0000-0002-6582-0275 (2022) RIPpore: A Novel Host-Derived Method for the Identification of Ricin Intoxication through Oxford Nanopore Direct RNA Sequencing. TOXINS, 14 (7). 470-.

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Official URL: http://dx.doi.org/10.3390/toxins14070470

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.

Item Type:	Article
Uncontrolled Keywords:	MinIon, ribosome inactivating proteins, ribosomes, ricin, RNA sequencing, saporin
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Infection, Veterinary and Ecological Sciences
Depositing User:	Symplectic Admin
Date Deposited:	18 Oct 2022 09:54
Last Modified:	18 Jan 2023 20:23
DOI:	10.3390/toxins14070470
Open Access URL:	https://doi.org/10.3390/toxins14070470
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3165589