{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T10:24:48Z","timestamp":1790763888163,"version":"4.1.0"},"posted":{"date-parts":[[2023,9,29]]},"group-title":"Microbiology","reference-count":52,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,9,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Rift Valley fever phlebovirus (RVFV) is a zoonotic pathogen that causes Rift Valley fever (RVF) in livestock and humans. Currently, there is no licensed human vaccine or antiviral drug to control RVF. Although multiple species of animals and humans are vulnerable to RVFV infection, host factors affecting susceptibility are not well understood.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methodology<\/jats:title>\n                  <jats:p>\n                    To identify the host factors or genes essential for RVFV replication, we conducted a CRISPR-Cas9 knock-out screen in human A549 cells. We then validated the putative genes using siRNA-mediated knockdowns and CRISPR-Cas9-mediated knockout studies, respectively. The role of a candidate gene in the virus replication cycle was assessed by measuring intracellular viral RNA accumulation, and the virus titers by plaque assay or TCID\n                    <jats:sub>50<\/jats:sub>\n                    assay.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Findings<\/jats:title>\n                  <jats:p>\n                    We identified approximately 900 genes with potential involvement in RVFV infection and replication. Further evaluation of the effect of six genes on viral replication using siRNA-mediated knockdowns found that silencing two genes (WDR7 and LRP1) significantly impaired RVFV replication. For further analysis, we focused on the\n                    <jats:italic>WDR7<\/jats:italic>\n                    gene since the role of\n                    <jats:italic>LRP1<\/jats:italic>\n                    in RVFV replication was previously described in detail. Knock-out A549 cell lines were generated and used to dissect the effect of\n                    <jats:italic>WRD7<\/jats:italic>\n                    on RVFV and another bunyavirus, La Crosse encephalitis virus (LACV). We observed significant effects of\n                    <jats:italic>WDR7<\/jats:italic>\n                    knock-out cells on both intracellular RVFV RNA levels and viral titers. At the intracellular RNA level,\n                    <jats:italic>WRD7<\/jats:italic>\n                    affected RVFV replication at a later phase of its replication cycle (24h) when compared to LACV which was affected an earlier replication phase (12h).\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>\n                    In summary, we have identified\n                    <jats:italic>WDR7<\/jats:italic>\n                    as an essential host factor for the replication of two relevant bunyaviruses, RVFV and LACV. Future studies will investigate the mechanistic role by which\n                    <jats:italic>WDR7<\/jats:italic>\n                    facilitates Phlebovirus replication.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Authors Summary<\/jats:title>\n                  <jats:p>Rift Valley fever phlebovirus is a high consequence pathogen that infects multiple animal species and also humans. Currently, there are no control measures available to treat RVF in humans and to prevent the incursion of Rift Valley fever virus into non-endemic countries. RVFV poses a significant threat to animal and human health in countries where it is endemic. RVFV replication depends on the host\u2019s machinery to complete its replication cycle. Therefore, one way to control virus replication is to disrupt the interaction between the virus and the host proteins important for replication. In this study, we identified a host factor, the WDR7 gene, that is critical for RVFV replication. The identification of this host factor is important as it can potentially lead to the development of antiviral strategies to control Rift Valley fever in both humans and animals.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.09.28.559935","type":"posted-content","created":{"date-parts":[[2023,9,29]],"date-time":"2023-09-29T20:55:14Z","timestamp":1696020914000},"source":"Crossref","is-referenced-by-count":2,"title":["Identification of host factors for Rift Valley Fever Phlebovirus"],"prefix":"10.64898","author":[{"given":"Velmurugan","family":"Balaraman","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabarish V.","family":"Indran","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yonghai","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David A.","family":"Meekins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laxmi U.M.R.","family":"Jakkula","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heidi","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Micheal P.","family":"Hays","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jayme A.","family":"Souza-Neto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natasha N.","family":"Gaudreault","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip R.","family":"Hardwidge","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William C.","family":"Wilson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Friedemann","family":"Weber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juergen A.","family":"Richt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2023100412150674000_2023.09.28.559935v1.1","doi-asserted-by":"publisher","DOI":"10.1002\/path.1700340418"},{"key":"2023100412150674000_2023.09.28.559935v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/0923-2516(91)90070-J"},{"key":"2023100412150674000_2023.09.28.559935v1.3","doi-asserted-by":"publisher","DOI":"10.3201\/eid0812.020195"},{"key":"2023100412150674000_2023.09.28.559935v1.4","doi-asserted-by":"publisher","DOI":"10.3201\/eid1504.081045"},{"key":"2023100412150674000_2023.09.28.559935v1.5","unstructured":"Coetzer JA . Natural Cases in New-Born Lambs. 1977;44: 205\u2013212."},{"key":"2023100412150674000_2023.09.28.559935v1.6","first-page":"11","article-title":"The pathology of Rift Valley fever. II. Lesions occurring in field cases in adult cattle, calves and aborted foetuses","volume":"49","year":"1982","journal-title":". Onderstepoort J Vet Res"},{"key":"2023100412150674000_2023.09.28.559935v1.7","doi-asserted-by":"publisher","DOI":"10.1051\/vetres\/2010033"},{"key":"2023100412150674000_2023.09.28.559935v1.8","doi-asserted-by":"publisher","DOI":"10.3201\/eid2409.180265"},{"key":"2023100412150674000_2023.09.28.559935v1.9","doi-asserted-by":"publisher","DOI":"10.1093\/ofid\/ofw200"},{"key":"2023100412150674000_2023.09.28.559935v1.10","doi-asserted-by":"publisher","DOI":"10.3390\/v3050493"},{"key":"2023100412150674000_2023.09.28.559935v1.11","doi-asserted-by":"publisher","DOI":"10.1017\/S0022172400062434"},{"key":"2023100412150674000_2023.09.28.559935v1.12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001421"},{"key":"2023100412150674000_2023.09.28.559935v1.13","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02341-13"},{"key":"2023100412150674000_2023.09.28.559935v1.14","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0131080"},{"key":"2023100412150674000_2023.09.28.559935v1.15","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2015.00001"},{"key":"2023100412150674000_2023.09.28.559935v1.16","doi-asserted-by":"publisher","DOI":"10.1177\/0300985819882633"},{"key":"2023100412150674000_2023.09.28.559935v1.17","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens10060681"},{"key":"2023100412150674000_2023.09.28.559935v1.18","doi-asserted-by":"publisher","DOI":"10.1099\/jgv.0.000765"},{"key":"2023100412150674000_2023.09.28.559935v1.19","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-81132-8"},{"key":"2023100412150674000_2023.09.28.559935v1.20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0002816"},{"key":"2023100412150674000_2023.09.28.559935v1.21","doi-asserted-by":"publisher","DOI":"10.2478\/jvetres-2021-0009"},{"key":"2023100412150674000_2023.09.28.559935v1.22","doi-asserted-by":"publisher","DOI":"10.1603\/ME10007"},{"key":"2023100412150674000_2023.09.28.559935v1.23","doi-asserted-by":"publisher","DOI":"10.1111\/mve.12254"},{"key":"2023100412150674000_2023.09.28.559935v1.24","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.13206"},{"key":"2023100412150674000_2023.09.28.559935v1.25","doi-asserted-by":"publisher","DOI":"10.1128\/msystems.01323-20"},{"key":"2023100412150674000_2023.09.28.559935v1.26","doi-asserted-by":"publisher","DOI":"10.1016\/0882-4010(87)90126-4"},{"key":"2023100412150674000_2023.09.28.559935v1.27","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00016-16"},{"key":"2023100412150674000_2023.09.28.559935v1.28","doi-asserted-by":"publisher","DOI":"10.1016\/j.physbeh.2017.03.040"},{"key":"2023100412150674000_2023.09.28.559935v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.09.001"},{"key":"2023100412150674000_2023.09.28.559935v1.30","doi-asserted-by":"publisher","DOI":"10.1111\/tra.12393"},{"key":"2023100412150674000_2023.09.28.559935v1.31","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2011.06.007"},{"key":"2023100412150674000_2023.09.28.559935v1.32","doi-asserted-by":"publisher","DOI":"10.3390\/v8010022"},{"key":"2023100412150674000_2023.09.28.559935v1.33","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02055-15"},{"key":"2023100412150674000_2023.09.28.559935v1.34","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01364-12"},{"key":"2023100412150674000_2023.09.28.559935v1.35","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02628-12"},{"key":"2023100412150674000_2023.09.28.559935v1.36","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2021.12.010"},{"key":"2023100412150674000_2023.09.28.559935v1.37","doi-asserted-by":"publisher","DOI":"10.26508\/lsa.202302005"},{"key":"2023100412150674000_2023.09.28.559935v1.38","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-66-10-2271"},{"key":"2023100412150674000_2023.09.28.559935v1.39","doi-asserted-by":"publisher","DOI":"10.1128\/genomeA.00926-16"},{"key":"2023100412150674000_2023.09.28.559935v1.40","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3047"},{"key":"2023100412150674000_2023.09.28.559935v1.41","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2017.016"},{"key":"2023100412150674000_2023.09.28.559935v1.42","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-018-0113-7"},{"key":"2023100412150674000_2023.09.28.559935v1.43","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.01234"},{"key":"2023100412150674000_2023.09.28.559935v1.44","doi-asserted-by":"publisher","DOI":"10.3892\/mmr.2015.3159"},{"key":"2023100412150674000_2023.09.28.559935v1.45","unstructured":"Haimes J , Kelley M . Demonstration of a \u0394\u0394Cq calculation method to compute relative gene expression from qPCR Data. Josh. GE Healthc. 2010. Available: http:\/\/dharmacon.gelifesciences.com\/uploadedfiles\/resources\/delta-cq-solaris-technote.pdf."},{"key":"2023100412150674000_2023.09.28.559935v1.46","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2017.01.015"},{"key":"2023100412150674000_2023.09.28.559935v1.47","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2008.12.011"},{"key":"2023100412150674000_2023.09.28.559935v1.48","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1262"},{"key":"2023100412150674000_2023.09.28.559935v1.49","doi-asserted-by":"publisher","DOI":"10.1016\/b978-012465330-6\/50003-8"},{"key":"2023100412150674000_2023.09.28.559935v1.50","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2016.06.028"},{"key":"2023100412150674000_2023.09.28.559935v1.51","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-13965-x"},{"key":"2023100412150674000_2023.09.28.559935v1.52","doi-asserted-by":"publisher","DOI":"10.1038\/srep08026"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2023.09.28.559935","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T05:18:06Z","timestamp":1768454286000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2023.09.28.559935"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,29]]},"references-count":52,"URL":"https:\/\/doi.org\/10.1101\/2023.09.28.559935","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.3390\/v15112251","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2023,9,29]]},"subtype":"preprint"}}