{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T12:12:46Z","timestamp":1792843966639,"version":"4.5.0"},"reference-count":61,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,5,11]],"date-time":"2021-05-11T00:00:00Z","timestamp":1620691200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001323","name":"Poliomyelitis Research Foundation","doi-asserted-by":"publisher","award":["13\/52"],"award-info":[{"award-number":["13\/52"]}],"id":[{"id":"10.13039\/501100001323","id-type":"DOI","asserted-by":"publisher"}]},{"name":"University of Pretoria, Institutional Research Theme","award":["Animal and Zoonotic Diseases (AZD)"],"award-info":[{"award-number":["Animal and Zoonotic Diseases (AZD)"]}]},{"name":"National Research Foundation South Africa","award":["97655"],"award-info":[{"award-number":["97655"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>If the goal of eliminating dog-mediated human rabies by 2030 is to be achieved, effective mass dog vaccination needs to be complemented by effective prophylaxis for individuals exposed to rabies. Aptamers and short-interfering RNAs (siRNAs) have been successful in therapeutics, but few studies have investigated their potential as rabies therapeutics. In this study, siRNAs and aptamers\u2014using a novel selection method\u2014were developed and tested against rabies virus (RABV) in a post-infection (p.i.) scenario. Multiple means of delivery were tested for siRNAs, including the use of Lipofectamine and conjugation with the developed aptamers. One siRNA (N53) resulted in an 80.13% reduction in viral RNA, while aptamer UPRET 2.03 demonstrated a 61.3% reduction when used alone at 2 h p.i. At 24 h p.i., chimera UPRET 2.03-N8 (aptamer-siRNA) resulted in a 36.5% inhibition of viral replication. To our knowledge, this is the first study using siRNAs or aptamers that (1) demonstrated significant inhibition of RABV using an aptamer, (2) tested Lipofectamine RNAi-Max as a means for delivery, and (3) produced significant RABV inhibition at 24 h p.i. This study serves as a proof-of-concept to potentially use aptamers and siRNAs as rabies immunoglobulin (RIG) replacements or therapeutic options for RABV and provides strong evidence towards their further investigation.<\/jats:p>","DOI":"10.3390\/v13050881","type":"journal-article","created":{"date-parts":[[2021,5,11]],"date-time":"2021-05-11T22:53:40Z","timestamp":1620773620000},"page":"881","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Rabies Prophylactic and Treatment Options: An In Vitro Study of siRNA- and Aptamer-Based Therapeutics"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5027-0031","authenticated-orcid":false,"given":"Terence Peter","family":"Scott","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Genetics and Microbiology, University of Pretoria, Pretoria 0002, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8963-0153","authenticated-orcid":false,"given":"Louis Hendrik","family":"Nel","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Genetics and Microbiology, University of Pretoria, Pretoria 0002, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e828","DOI":"10.1016\/S2214-109X(18)30302-4","article-title":"New global strategic plan to eliminate dog-mediated rabies by 2030","volume":"6","author":"Minghui","year":"2018","journal-title":"Lancet Glob. Health"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Wallace, R.M., Undurraga, E.A., Blanton, J.D., Cleaton, J., and Franka, R. (2017). Elimination of Dog-mediated human rabies deaths by 2030: Needs assessmnet and alternatives for progress based on dog vaccination. Front. Vet. Sci., 4.","DOI":"10.3389\/fvets.2017.00009"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/j.antiviral.2013.03.011","article-title":"Eliminating canine rabies, the principal source of human infection: What will it take?","volume":"98","author":"Meslin","year":"2013","journal-title":"Antivir. Res."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Hampson, K., Coudeville, L., Lembo, T., Sambo, M., Kieffer, A., Attlan, M., Barrat, J., Blanton, J.D., Briggs, D.J., and Cleaveland, S. (2015). Estimating the Global Burden of Endemic Canine Rabies. PLoS Negl. Trop. Dis., 9.","DOI":"10.1371\/journal.pntd.0003786"},{"key":"ref_5","unstructured":"World Health Organisation (2018). WHO Expert Consultation on Rabies, World Health Organization. [3rd ed.]."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"14996","DOI":"10.1073\/pnas.0904740106","article-title":"Transmission dynamics and economics of rabies control in dogs and humans in an African city","volume":"106","author":"Zinsstag","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Hampson, K., Cleaveland, S., and Briggs, D. (2011). Evaluation of cost-effective strategies for rabies post-exposure vaccination in low-income countries. PLoS Negl. Trop. Dis., 5.","DOI":"10.1371\/journal.pntd.0000982"},{"key":"ref_8","first-page":"360","article-title":"Re-evaluating the burden of rabies in Africa and Asia","volume":"83","author":"Knobel","year":"2005","journal-title":"Bull. World Health Organ."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"M\u00fcller, T., Dietzschold, B., Ertl, H., Fooks, A.R., Freuling, C., Fehlner-Gardiner, C., Kliemt, J., Meslin, F.X., Rupprecht, C.E., and Tordo, N. (2009). Development of a mouse monoclonal antibody cocktail for post-exposure rabies prophylaxis in humans. PLoS Negl. Trop. Dis., 3.","DOI":"10.1371\/annotation\/df98339d-6bdb-40ed-af83-cc38b249264a"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"407","DOI":"10.15252\/emmm.201505986","article-title":"Development of broad-spectrum human monoclonal antibodies for rabies post-exposure prophylaxis","volume":"8","author":"Minola","year":"2016","journal-title":"EMBO Mol. Med."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Kim, P.K., Keum, S.J., Osinubi, M.O.V., Franka, R., Shin, J.Y., Park, S.T., Kim, M.S., Park, M.J., Lee, S.Y., and Carson, W. (2017). Development and characterization of novel chimeric monoclonal antibodies for broad spectrum neutralization of rabies virus. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0186380"},{"key":"ref_12","unstructured":"Strategic Advisory Group of Experts on Immunization, and World Health Organisation (2017). Background Paper: Proposed Revision of the Policy on Rabies Vaccines and Rabies Immunoglobulins, World Health Organization."},{"key":"ref_13","unstructured":"World Health Organization (2013). WHO Expert Consultation on Rabies, World Health Organization. [2nd ed.]. World Health Organization Tech. Rep. Ser."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2508","DOI":"10.1056\/NEJMoa050382","article-title":"Survival after Treatment of Rabies with Induction of Coma","volume":"352","author":"Willoughby","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_15","unstructured":"(2018, January 26). Medical College of Wisconsin Rabies Registry. Available online: http:\/\/www.mcw.edu\/Pediatrics\/InfectiousDiseases\/PatientCare\/Rabies.htm."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1017\/cjn.2015.331","article-title":"Critical appraisal of the Milwaukee protocol for rabies: This failed approach should be abandoned","volume":"43","author":"Zeiler","year":"2016","journal-title":"Can. J. Neurol. Sci. J. Can. Sci. Neurol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1097\/INF.0000000000000641","article-title":"The \u201cMilwaukee Protocol\u201d for Treatment of Human Rabies Is No Longer Valid","volume":"34","author":"Wilde","year":"2015","journal-title":"Pediatr. Infect. Dis. J."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Scott, T.P., and Nel, L.H. (2016). Subversion of the Immune Response by Rabies Virus. Viruses, 8.","DOI":"10.3390\/v8080231"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"260","DOI":"10.15302\/J-FASE-2016116","article-title":"Critical roles of chemokines and cytokines in antiviral innate immune responses during rabies virus infection","volume":"4","author":"Huang","year":"2017","journal-title":"Front. Agric. Sci. Eng."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3200","DOI":"10.1128\/JVI.06699-11","article-title":"Postexposure treatment with the live-attenuated rabies virus (RV) vaccine TriGAS triggers the clearance of wild-type RV from the Central Nervous System (CNS) through the rapid induction of genes relevant to adaptive immunity in CNS tissues","volume":"86","author":"Li","year":"2012","journal-title":"J. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1038\/35078107","article-title":"Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells","volume":"411","author":"Elbashir","year":"2001","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1038\/35005107","article-title":"An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells","volume":"404","author":"Hammond","year":"2000","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/S0092-8674(00)80620-0","article-title":"RNAi: Double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals","volume":"101","author":"Zamore","year":"2000","journal-title":"Cell"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1038\/35053110","article-title":"Role for a bidentate ribonuclease in the initiation step of RNA interference","volume":"409","author":"Bernstein","year":"2001","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/j.cell.2005.08.044","article-title":"Passenger-strand cleavage facilitates assembly of siRNA into Ago2-containing RNAi enzyme complexes","volume":"123","author":"Matranga","year":"2005","journal-title":"Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6877","DOI":"10.1093\/emboj\/20.23.6877","article-title":"Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate","volume":"20","author":"Elbashir","year":"2001","journal-title":"EMBO J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1101\/gad.862301","article-title":"RNA interference is mediated by 21- and 22-nucleotide RNAs","volume":"15","author":"Elbashir","year":"2001","journal-title":"Genes Dev."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1093\/nar\/gkj439","article-title":"Insights into the kinetics of siRNA-mediated gene silencing from live-cell and live-animal bioluminescent imaging","volume":"34","author":"Bartlett","year":"2006","journal-title":"Nucleic Acids Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1002\/bit.21285","article-title":"Effect of siRNA Nuclease Stability on the In vitro and in vivo kinetics of siRNA-mediated Gene Silencing","volume":"97","author":"Bartlett","year":"2007","journal-title":"Biotechnol. Bioeng."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1000","DOI":"10.1016\/S0006-291X(02)02013-2","article-title":"Comparison of antisense oligonucleotides and siRNAs in cell culture and in vivo","volume":"296","author":"Bertrand","year":"2002","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.addr.2007.03.005","article-title":"RNAi therapeutics: Principles, prospects and challenges","volume":"59","author":"Aagaard","year":"2007","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1186\/s13073-017-0450-0","article-title":"Advances in the delivery of RNA therapeutics: From concept to clinical reality","volume":"9","author":"Kaczmarek","year":"2017","journal-title":"Genome Med."},{"key":"ref_33","first-page":"5","article-title":"RNAi\u2014Based Treatment for Rabies: Current Achievements and Insights Towards a Cure","volume":"2","year":"2015","journal-title":"Br. J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"566","DOI":"10.1016\/j.bjm.2016.11.007","article-title":"Short interfering RNAs targeting a vampire-bat related rabies virus phosphoprotein mRNA","volume":"48","author":"Ono","year":"2017","journal-title":"Braz. J. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"29","DOI":"10.3389\/fmicb.2012.00029","article-title":"Aptamers in virology: Recent advances and challenges","volume":"3","author":"Binning","year":"2012","journal-title":"Front. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"612","DOI":"10.3390\/s120100612","article-title":"Aptamers and their biological applications","volume":"12","author":"Song","year":"2012","journal-title":"Sensors"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1038\/346818a0","article-title":"In vitro selection of RNA molecules that bind specific ligands","volume":"346","author":"Ellington","year":"1990","journal-title":"Nature"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1126\/science.2200121","article-title":"Systematic Evolution of Ligands by Exponential Enrichment: RNA Ligands to Bacteriophage T4 DNA Polymerase","volume":"249","author":"Tuerk","year":"1990","journal-title":"Science"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1016\/0022-2836(91)90509-5","article-title":"SELEXION Systematic Evolution of Ligands by Exponential Enrichment with Integrated Optimization by Non-linear Analysis","volume":"222","author":"Irvine","year":"1991","journal-title":"J. Mol. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1146\/annurev-pharmtox-010716-104558","article-title":"Aptamers as therapeutics","volume":"57","author":"Nimjee","year":"2017","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Gonz\u00e1lez, V.M., Elena Mart\u00edn, M., Fern\u00e1ndez, G., and Garc\u00eda-Sacrist\u00e1n, A. (2016). Use of aptamers as diagnostics tools and antiviral agents for human viruses. Pharmaceuticals, 9.","DOI":"10.3390\/ph9040078"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e71","DOI":"10.1038\/mtna.2012.62","article-title":"Potent inhibition of HIV-1 reverse transcriptase and replication by nonpseudoknot, \u201cUCAA-motif\u201d RNA aptamers","volume":"2","author":"Whatley","year":"2013","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Larramendy, M.L., and Soloneski, S. (2016). Application of Nucleic Acid Aptamers to Viral Detection and Inhibition. Nucleic Acids\u2014From Basic Aspects to Laboratory Tools, InTech. Chapter 6.","DOI":"10.5772\/60691"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1089\/154545703322860771","article-title":"Selection of DNA aptamers that bind the RNA-dependent RNA polymerase of hepatitis C virus and inhibit viral RNA synthesis in vitro","volume":"13","author":"Bellecave","year":"2003","journal-title":"Oligonucleotides"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1007\/s10126-012-9448-1","article-title":"RNA aptamers inhibit the growth of the fish pathogen viral hemorrhagic septicemia virus (VHSV)","volume":"14","author":"Punnarak","year":"2012","journal-title":"Mar. Biotechnol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1111\/jfd.12000","article-title":"Inhibition of hirame rhabdovirus growth by RNA aptamers","volume":"35","author":"Hwang","year":"2012","journal-title":"J. Fish Dis."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/j.intimp.2012.06.019","article-title":"Isolation of ssDNA aptamers that inhibit rabies virus","volume":"14","author":"Liang","year":"2012","journal-title":"Int. Immunopharmacol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1016\/j.virusres.2012.12.017","article-title":"Aptamers targeting rabies virus-infected cells inhibit viral replication both in vitro and in vivo","volume":"173","author":"Liang","year":"2013","journal-title":"Virus Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"432","DOI":"10.1016\/j.intimp.2014.03.020","article-title":"Aptamers targeting rabies virus-infected cells inhibit street rabies virus in vivo","volume":"21","author":"Liang","year":"2014","journal-title":"Int. Immunopharmacol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.virusres.2014.01.021","article-title":"Selection of an aptamer against rabies virus: A new class of molecules with antiviral activity","volume":"184","author":"Liang","year":"2014","journal-title":"Virus Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"7261","DOI":"10.1128\/JVI.74.16.7261-7269.2000","article-title":"Differential transcription attenuation of rabies virus genes by intergenic regions: Generation of recombinant viruses overexpressing the polymerase gene","volume":"74","author":"Finke","year":"2000","journal-title":"J. Virol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"3949","DOI":"10.1128\/JCM.01256-10","article-title":"Improved PCR methods for detection of African rabies and rabies-related lyssaviruses","volume":"48","author":"Coertse","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"e73","DOI":"10.1093\/nar\/gkl388","article-title":"Aptamer mediated siRNA delivery","volume":"34","author":"Chu","year":"2006","journal-title":"Nucleic Acids Res."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.antiviral.2016.09.011","article-title":"The SARE tool for rabies control: Current experience in Ethiopia","volume":"135","author":"Coetzer","year":"2016","journal-title":"Antivir. Res."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"4","DOI":"10.3389\/fvets.2017.00018","article-title":"Addressing the disconnect between the estimated, reported and true rabies data: The development of a regional African rabies bulletin","volume":"4","author":"Scott","year":"2017","journal-title":"Front. Vet. Sci."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.antiviral.2015.10.002","article-title":"The Pan-African Rabies Control Network (PARACON): A unified approach to eliminating canine rabies in Africa","volume":"124","author":"Scott","year":"2015","journal-title":"Antivir. Res."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.antiviral.2018.07.018","article-title":"Formation of the Asian Rabies Control Network (ARACON): A common approach towards a global good","volume":"157","author":"Coetzer","year":"2018","journal-title":"Antivir. Res."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Gomme, E.A., Wirblich, C., Addya, S., Rall, G.F., and Schnell, M.J. (2012). Immune clearance of attenuated rabies virus results in neuronal survival with altered gene expression. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002971"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1007\/s00005-013-0227-0","article-title":"Application of aptamers for targeted therapeutics","volume":"61","author":"Ray","year":"2013","journal-title":"Arch. Immunol. Ther. Exp. (Warsz.)"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1021\/bc800278e","article-title":"siRNA Conjugate Delivery Systems","volume":"20","author":"Jeong","year":"2009","journal-title":"Bioconjug. Chem."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1399","DOI":"10.1038\/nbt1161","article-title":"Activation of the mammalian immune system by siRNAs","volume":"23","author":"Marques","year":"2005","journal-title":"Nat. Biotechnol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/5\/881\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:59:20Z","timestamp":1760162360000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/5\/881"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,11]]},"references-count":61,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["v13050881"],"URL":"https:\/\/doi.org\/10.3390\/v13050881","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,11]]}}}