{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T19:27:06Z","timestamp":1792178826371,"version":"4.4.0"},"reference-count":65,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T00:00:00Z","timestamp":1747699200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T00:00:00Z","timestamp":1747699200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100006502","name":"Defense Sciences Office, DARPA","doi-asserted-by":"publisher","award":["D18AC00028"],"award-info":[{"award-number":["D18AC00028"]}],"id":[{"id":"10.13039\/100006502","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006492","name":"Division of Intramural Research of the NIAID","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100006492","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Virol"],"published-print":{"date-parts":[[2025,5,20]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Herpesvirus-based vectors are attractive for use as conventional or transmissible vaccines against emerging zoonoses in inaccessible animal populations. In both cases, cytomegaloviruses (CMVs) as members of the subfamily\n              <jats:italic toggle=\"yes\">Betaherpesvirinae<\/jats:italic>\n              are particularly suitable for vaccine development as they are highly specific for their natural host species, infect a large proportion of their host population, and cause mild infections in healthy individuals. The Natal multimammate mouse (\n              <jats:italic toggle=\"yes\">Mastomys natalensis<\/jats:italic>\n              ) is the natural reservoir of Lassa virus, which causes deadly hemorrhagic fever in humans.\n              <jats:italic toggle=\"yes\">M. natalensis<\/jats:italic>\n              was recently reported to harbor at least three different cytomegaloviruses (MnatCMV1, MnatCMV2, and MnatCMV3). Herein, we report the molecular cloning of three complete MnatCMV genomes in a yeast and bacterial artificial chromosome (YAC-BAC) hybrid vector. Purified viral genomes were cloned in yeast by single-step transformation-associated recombination (STAR cloning) and subsequently transferred to\n              <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n              for further genetic manipulation. The integrity of the complete cloned viral genomes was verified by sequencing, and the replication fitness of viruses reconstituted from these clones was analyzed by replication kinetics in\n              <jats:italic toggle=\"yes\">M. natalensis<\/jats:italic>\n              fibroblasts and kidney epithelial cells. We also found that neither parental nor cloned MnatCMVs replicated in mouse and rat fibroblasts, nor did they show sustained replication in baby hamster kidney cells, consistent with the expected narrow host range for these viruses. We further demonstrated that an exogenous sequence can be inserted by BAC-based mutagenesis between open reading frames M25 and m25.1 of MnatCMV2 without affecting replication fitness\n              <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n              , identifying this site as potentially suitable for the insertion of vaccine target antigen genes.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                Cytomegaloviruses (CMVs) recently discovered in the Natal multimammate mouse (\n                <jats:italic toggle=\"yes\">Mastomys natalensis<\/jats:italic>\n                ) are widespread within the\n                <jats:italic toggle=\"yes\">M. natalensis<\/jats:italic>\n                population. Since these rodents also serve as natural hosts of the human pathogen Lassa virus (LASV), we investigated the potential suitability of\n                <jats:italic toggle=\"yes\">M. natalensis<\/jats:italic>\n                CMVs (MnatCMVs) as vaccine vectors. We describe the cloning of three different MnatCMV genomes as bacterial artificial chromosomes (BACs). The replicative capacity and species specificity of these BAC-derived MnatCMVs were analyzed in multiple cell types. We also identified a transgene insertion site within one of the MnatCMV genomes suitable for the incorporation of vaccine target antigens. Together, this study provides a foundation for the development of MnatCMVs as transmissible MnatCMV-based LASV vaccines to reduce LASV prevalence in hard-to-reach\n                <jats:italic toggle=\"yes\">M. natalensis<\/jats:italic>\n                populations and, thereby, zoonotic transmission to humans.\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/jvi.02147-24","type":"journal-article","created":{"date-parts":[[2025,4,9]],"date-time":"2025-04-09T13:05:02Z","timestamp":1744203902000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":4,"title":["Molecular cloning and host range analysis of three cytomegaloviruses from\n            <i>Mastomys natalensis<\/i>"],"prefix":"10.1128","volume":"99","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-5609-7643","authenticated-orcid":true,"given":"Laura","family":"Staliunaite","sequence":"first","affiliation":[{"name":"Leibniz Institute of Virology (LIV)","place":["Hamburg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5789-8988","authenticated-orcid":false,"given":"Olha","family":"Puhach","sequence":"additional","affiliation":[{"name":"Leibniz Institute of Virology (LIV)","place":["Hamburg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7539-2155","authenticated-orcid":true,"given":"Eleonore","family":"Ostermann","sequence":"additional","affiliation":[{"name":"Leibniz Institute of Virology (LIV)","place":["Hamburg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyle","family":"Rosenke","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health","place":["Hamilton, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0093-7170","authenticated-orcid":false,"given":"Jenna","family":"Nichols","sequence":"additional","affiliation":[{"name":"MRC-University of Glasgow Centre for Virus Research","place":["Glasgow, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0098-7485","authenticated-orcid":false,"given":"Lisa","family":"Oestereich","sequence":"additional","affiliation":[{"name":"Bernhard Nocht Institute for Tropical Medicine (BNITM)","place":["Hamburg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nafomon","family":"Sogoba","sequence":"additional","affiliation":[{"name":"University of Sciences, Techniques and Technologies of Bamako","place":["Bamako, Mali"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9448-8227","authenticated-orcid":true,"given":"Heinz","family":"Feldmann","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health","place":["Hamilton, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4991-9128","authenticated-orcid":true,"given":"Andrew J.","family":"Davison","sequence":"additional","affiliation":[{"name":"MRC-University of Glasgow Centre for Virus Research","place":["Glasgow, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0124-4061","authenticated-orcid":true,"given":"Michael A.","family":"Jarvis","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health","place":["Hamilton, USA"]},{"name":"The Vaccine Group Ltd.","place":["Plymouth, United Kingdom"]},{"name":"School of Biomedical Sciences, University of Plymouth","place":["Plymouth, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6078-5255","authenticated-orcid":false,"given":"Wolfram","family":"Brune","sequence":"additional","affiliation":[{"name":"Leibniz Institute of Virology (LIV)","place":["Hamburg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","first-page":"212","volume-title":"Fields Virology","author":"Krug LT","year":"2021","unstructured":"Krug LT, Pellett PE. 2021. 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Caister Academic Press, Norfolk, UK."},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-62703-788-4_15"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-020-04738-9"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1099\/jgv.0.001873"},{"key":"e_1_3_4_8_2","unstructured":"European Centre for Disease Prevention and Control. 2016. Rapid risk assessment. Lassa fever in Nigeria Benin Togo Germany and USA European centre for disease prevention and control (ECDC) ECfDPaC. Stockholm."},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1804498"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1111\/2041-210X.12549"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2688"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines7040152"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.3390\/v15102043"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001275"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2015.03.029"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1135(88)90043-0"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1185350"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1155\/2011\/124595"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1155\/2012\/472537"},{"key":"e_1_3_4_20_2","first-page":"38","volume-title":"Cytomegaloviruses: from molecular pathogenesis to intervention","author":"Ruzsics Z","year":"2013","unstructured":"Ruzsics Z, Borst EM, Bosse JB, Brune W, Messerle M. 2013. Manipulating CMV genomes by BAC mutagenesis: strategies and applications, p 38\u201358. In Reddehase MJ (ed), Cytomegaloviruses: from molecular pathogenesis to intervention. Caister Academic Press, Norfolk, UK."},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.26.14759"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1700534114"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/mSphereDirect.00331-17"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.crmeth.2024.100696"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.1.48-56.1998"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1093\/femsre\/fuy004"},{"key":"e_1_3_4_27_2","first-page":"420","article-title":"Murine virus susceptibility of cell cultures of mouse, rat, hamster, monkey, and human origin","volume":"25","author":"Reed JM","year":"1975","unstructured":"Reed JM, Schiff LJ, Shefner AM, Poiley SM. 1975. Murine virus susceptibility of cell cultures of mouse, rat, hamster, monkey, and human origin. 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Adaptation of a cytomegalic virus from fieldmice (Apodemus sylvaticus) on various heterologous cells, human included. C R Acad Hebd Seances Acad Sci D 269:104\u2013106.","journal-title":"C R Acad Hebd Seances Acad Sci D"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.25.8.1456-1458.1987"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-79-2-347"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.7.6.720-725.1971"},{"key":"e_1_3_4_38_2","article-title":"Isolation of five strains of herpesviruses from two species of free living small rodents","volume":"24","author":"Blaskovic D","year":"1980","unstructured":"Blaskovic D, Stancekov\u00e1 M, Svobodov\u00e1 J, Mistr\u00edkov\u00e1 J. 1980. Isolation of five strains of herpesviruses from two species of free living small rodents. 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