Journal of General Virology最新文献

{"title":"Update on antimicrobial peptides: key elements in <i>Orthoflavivirus</i> infection - an overview.","authors":"Victor Javier Cruz-Holguín, Ivonne Sagrario Romero-Flores, Luis Gerardo Olmos-Bustos, Julio García-Cordero, Moisés León-Juárez, Leticia Cedillo-Barron","doi":"10.1099/jgv.0.002129","DOIUrl":"10.1099/jgv.0.002129","url":null,"abstract":"<p><p><i>Flaviviridae</i> is a family of viruses that are mainly transmitted by mosquito vectors of the genus <i>Aedes</i>, which cause febrile illnesses and, in severe cases, haemorrhagic or neurodegenerative conditions. Over time, these viruses have been reported as emerging pathogens, leading to epidemic outbreaks in various regions worldwide. Additionally, climate change has facilitated the migration of these vectors to regions where they were not previously found. Unfortunately, there are currently no effective treatments or vaccines to prevent or combat <i>Orthoflavivirus</i> infections. Consequently, a deeper understanding of the viral biology and the human host immune response is crucial for advancing the development of therapeutic targets. Amongst the molecules involved in the innate immune response to viral infections are antimicrobial peptides (AMPs), which have been studied for decades. However, their role in <i>Orthoflavivirus</i> infections remains poorly understood. Several researchers have proposed the stimulation or exogenous administration of AMPs during various viral infections, highlighting these molecules as potential innovative therapeutic targets. This study compiles current knowledge on AMPs with a specific focus on <i>Orthoflavivirus</i> infections, emphasizing the importance of these promising therapeutic approaches.</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12225737/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144553745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Susceptibility and transmission of mpox virus infection in brown rats (<i>Rattus norvegicus</i>).","authors":"Lucy Crossley, Stephen Findlay-Wilson, Linda Easterbrook, Emma Kennedy, Francisco J Salguero, Kim Mackay, Victoria Graham, Susan Fotheringham, Stuart Dowall","doi":"10.1099/jgv.0.002125","DOIUrl":"10.1099/jgv.0.002125","url":null,"abstract":"<p><p>Mpox (formerly known as monkeypox) virus (MPXV) is the zoonotic pathogen of mpox disease in humans. Its increasing emergence outside of its endemic area has heightened the importance of investigating the virus' prevalence and maintenance in sylvatic reservoirs. The common brown rat (<i>Rattus norvegicus</i>) can inhabit almost anywhere in the UK, posing a threat to zoonotic transmission to humans. Two independent studies were carried out; the first investigated the susceptibility of brown rats to MPXV infection with a clade IIb mpox strain via two challenge routes: intranasal and intradermal. The second study considered the transmission of MPXV between challenged and naïve brown rats. All animals were asymptomatic to mpox disease, although enzyme-linked immunosorbent assay (ELISA) confirmed subclinical infection in challenge groups. In the susceptibility study, reverse transcription PCR (RT-PCR) detected mpox DNA in the lung tissue and throat swabs within the intranasally inoculated group, in addition to viable virus observed from the intranasal throat swabs. In contrast, no virus was detected in either tissues or swabs in the intradermally inoculated group or control group. RT-PCR results from the transmission study detected mpox DNA in tissues and throat swabs taken from challenged animals. Viable virus was observed from tissues and swabs of intranasally challenged animals with infectious titres of ~10²-10⁴ TCID₅₀ per millilitre. ELISA assays in the transmission study showed replicable results compared to the first susceptibility study in directly challenged animals alongside evidence of seroconversion in co-housed naïve animals. In conclusion, brown rats are susceptible to MPXV infection, as they have been demonstrated to maintain viable virus in the absence of clinical signs. Viral transmission of MPXV from infected rats to naïve rats was not observed by RT-PCR, although naïve rats did show antibody responses when exposed to infected rats indicating exposure to virus.</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12214244/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144540487","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Summary of taxonomy changes ratified by the International Committee on Taxonomy of Viruses (ICTV) from the Bacterial Viruses Subcommittee, 2025.","authors":"Dann Turner, Evelien M Adriaenssens, Rudolf I Amann, Pavol Bardy, Nina Bartlau, Jakub Barylski, Stanisław Błażejak, Majid Bouzari, Ariane Briegel, Yves Briers, Daniel Carrillo, Xia Chen, Dennis Claessen, Ryan Cook, Marco A Crisci, Arnaud Dechesne, Paulina Deptula, Bas E Dutilh, Bert Ely, Lars Fieseler, Paul C M Fogg, Akihito Fukudome, Mohammed Saeed Ganjoor, Iwona Gientka, Karin Holmfeldt, Panos G Kalatzis, Kathryn M Kauffman, Annabel Kempff, Petar Knezevic, Eugene V Koonin, Andrew M Kropinski, Mart Krupovic, Ipek Kurtböke, Kai Lambon, Rob Lavigne, Susan M Lehman, H-T Liu, Cedric Lood, Rudi Lurz, Sari Mäntynen, Cole B Matrishin, Mathias Middelboe, Andrew D Millard, Cristina Moraru, Dennis S Nielsen, Franklin L Nobrega, Takuro Nunoura, Hanna M Oksanen, Véronique Ongenae, Boris Parra, Célia Pas, Joseph Pogliano, Minna M Poranen, Siravudh Potipimpanon, Amy Prichard, Hannah V Pye, Daniela Rothschild-Rodriguez, Daniel E Rozen, Joanne M Santini, Yuandong Sha, Dziyana Shymialevich, Barbara Sokołowska, Abbas Soleimani-Delfan, Paulina Średnicka, Paulo Tavares, Andrea Telatin, Igor Tolstoy, Shyun-Ichi Urayama, Vera van Neer, Finn K Vogensen, Qiannan Wen, Antje Wichels, Michał Wójcicki, Ictv Taxonomy Summary Consortium","doi":"10.1099/jgv.0.002111","DOIUrl":"10.1099/jgv.0.002111","url":null,"abstract":"<p><p>This article summarises the activities of the International Committee on Taxonomy of Viruses Bacterial Viruses Subcommittee, detailing developments in the classification of bacterial viruses. We provide here an overview of all new, abolished, moved and renamed taxa proposed in 2024, approved by the Executive Committee, and ratified by membership vote in 2025. Through the collective efforts of 74 international contributors of taxonomy proposals in this round, 43 ratified proposals have led to the creation of one new phylum, one class, four orders, 33 families, 14 subfamilies, 194 genera and 995 species. These proposals mark significant progress in refining the taxonomy of bacterial viruses. Key updates include the creation of new orders and families that include existing taxa to better reflect genomic and evolutionary relationships. As sequencing and bioinformatics approaches continue to advance, further expansion and refinements in viral taxonomy can be anticipated in the coming years.</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144715088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Kinesin light chain 1 interacts with NS1 and is a susceptibility factor for dengue virus infection in mosquito cells.","authors":"Juan Manuel Castillo, Raymundo Cruz-Pérez, Daniel Talamás-Lara, Juan E Ludert","doi":"10.1099/jgv.0.002132","DOIUrl":"10.1099/jgv.0.002132","url":null,"abstract":"<p><p>A hallmark of the dengue virus (DENV) infection is the manipulation of host cell membranes, lipid trafficking and lipid droplets (LD), all cellular functions that depend on the cytoskeleton and the cytoplasmic streaming system. We previously reported the interaction between the DENV non-structural (NS1) protein and members of the kinesin motor complex in the <i>Aedes albopictus</i> cell line C6/36. In this work, we present evidence indicating that the protein kinesin light chain 1 (KLC1) is indeed a susceptibility factor for the DENV replicative cycle in mosquito cells. The interaction between NS1 and KLC1 was confirmed by proximity ligation and co-immunoprecipitation assays in cells harvested 24 hpi. In addition, transmission immunoelectron microscopy showed KLC1 decorating the surface of vacuoles in association with NS1. Increased levels of KLC1 were observed starting at 6 hpi, suggesting that virus infection stimulates KLC1 synthesis. Silencing KLC1 expression results in a reduction in viral genome synthesis, decreased secretion of NS1 and a reduction of virus progeny by nearly 1 log. In agreement, similar affectations were observed in infected cells transfected with a peptide that competes and interferes with the interaction between KLC1 and its cargo molecules. Of note, both silencing the expression and interfering with the function of KLC1 resulted in a disorganization of LD, which decreased in number and increased in area, in mock or infected cells. These results, taken together, suggest that KLC1 is a host susceptibility factor for DENV in mosquito cells and appears to play an important role in the proper transport and homeostasis of LD required for flavivirus replication. However, modest colocalization was observed between NS1 and LD, and the significance of the KLC1 and NS1 interactions needs to be further investigated.</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12266351/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144642752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of pandemic potential of the genotype 4 (G4) swine influenza virus using <i>ex vivo</i> and <i>in vitro</i> cultures of the human respiratory tract.","authors":"Jenny C M Chan, Rachel H H Ching, Hermione H M Kock, Teng Long, John M Nicholls, J S Malik Peiris, Kenrie P Y Hui, Michael C W Chan","doi":"10.1099/jgv.0.002133","DOIUrl":"10.1099/jgv.0.002133","url":null,"abstract":"<p><p>Recent studies have reported a genotype 4 (G4) reassortant Eurasian avian-like (EA) H1N1 virus in swine, demonstrating a potential pandemic threat in humans. Here, we have compared the tropism, replication competence and pro-inflammatory cytokine and chemokine induction of the two G4 EA H1N1 strains in parallel with 2009 pandemic H1N1 (H1N1/pdm/09) and A/Quail/HK/G1/1997 H9N2 (G1) using <i>ex vivo</i> culture of the human respiratory tract and <i>in vitro</i> culture of human peripheral blood-derived macrophages. Our results showed that G4 strains could replicate in <i>ex vivo</i> cultures of human lung and bronchus with a similar replication competence to H1N1/pdm/09. The cytokine induction levels of G4 were similar to H1N1/pdm/09 in macrophages. Taken together, we could extrapolate that the G4 EA H1N1 swine influenza may pose a notable public health threat towards human and should not underestimate this threat.</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12313140/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144760296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Summary of taxonomy changes ratified by the International Committee on Taxonomy of Viruses (ICTV) from the Fungal and Protist Viruses Subcommittee, 2025.","authors":"Sead Sabanadzovic, Chantal Abergel, Marı́a A Ayllón, Leticia Botella, Marta Canuti, Yuto Chiba, Jean-Michel Claverie, Robert H A Coutts, Stefania Daghino, Livia Donaire, Marco Forgia, Ondřej Hejna, Jichun Jia, Daohong Jiang, Ioly Kotta-Loizou, Mart Krupovic, Andrew S Lang, Matthieu Legendre, Shin-Yi Lee Marzano, Luca Nerva, Judit Pénzes, Anna Poimala, Sofia Rigou, Yukiyo Sato, Wajeeha Shamsi, Nobuhiro Suzuki, Massimo Turina, Syun-Ichi Urayama, Eeva J Vainio, Jiatao Xie, Ictv Taxonomy Summary Consortium","doi":"10.1099/jgv.0.002115","DOIUrl":"10.1099/jgv.0.002115","url":null,"abstract":"<p><p>The Fungal and Protist Viruses Subcommittee (SC) of the International Committee on Taxonomy of Viruses (ICTV) has received a total of eight taxonomic proposals for the 2024 annual cycle. The extent of proposed changes varied, including nomenclatural updates, creation of new taxa and reorganization of established taxa. Following the ICTV procedures, all proposals were reviewed and voted upon by the members of the Executive Committee with ratification in March 2025. As a result, a total of 52 species in the families <i>Botourmiaviridae</i> and <i>Marnaviridae</i> were renamed to comply with the mandated binomial format. A new genus has been added to the dsRNA virus family <i>Amalgaviridae</i>, while two new families, <i>Splipalmiviridae</i> (<i>Wolframvirales</i>) and <i>Mycoalphaviridae</i> (<i>Hepelivirales</i>), were created to classify new groups of positive-sense (+) RNA mycoviruses. The class <i>Arfiviricetes</i> (<i>Cressdnaviricota</i>) was expanded by a new order <i>Lineavirales</i> and a new family <i>Oomyviridae</i> of ssDNA viruses. Additionally, a new class <i>Orpoviricetes</i> was created in the kingdom <i>Orthornavirae</i> to classify a group of bisegmented (+)RNA viruses reported from fungi and oomycetes. Finally, the order <i>Pimascovirales</i> was reorganized to better depict evolutionary relationships of pithoviruses and related viruses with large dsDNA genomes. The summary of updates in the taxonomy of fungal and protist viruses presented here is limited to taxa within the remit of this Subcommittee. For information on taxonomy changes on other fungal viruses closely related to animal and/or plant viruses, please see reports from sister ICTV Subcommittees (i.e. Plant Virus SC and Animal dsRNA and ssRNA(-) Viruses SC).</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144707726","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Summary of taxonomy changes ratified by the International Committee on Taxonomy of Viruses (ICTV) - General taxonomy proposals, 2025.","authors":"F Murilo Zerbini, Anya Crane, Jens H Kuhn, Peter Simmonds, Elliot J Lefkowitz, Ictv Taxonomy Summary Consortium","doi":"10.1099/jgv.0.002116","DOIUrl":"10.1099/jgv.0.002116","url":null,"abstract":"<p><p>During the 56th annual meeting of the International Committee on Taxonomy of Viruses (ICTV), held in Bari, Italy, in August 2024, two technical proposals were presented. The first called for amended versions of accepted taxonomic proposals to be named in such a way to ensure that they are readily accessible on the ICTV website (2024.001G). The second proposed a substantial reformatting of the ICTV statutes and codes to produce a more unified text after the numerous changes made to both documents in previous years (2024.002G). Finally, the ICTV Executive Committee (EC) nominated Professor Stuart Siddell as a Life Member of the ICTV for his work over four decades on virus taxonomy, including 16 years as a member of the EC (2024.003G).</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144715086","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Understanding the mechanisms of mitochondrial rewiring during viral infections.","authors":"Marta Lopez-Nieto, Nicolas Locker","doi":"10.1099/jgv.0.002128","DOIUrl":"10.1099/jgv.0.002128","url":null,"abstract":"<p><p>As intracellular parasites, viruses must hijack and often rewire organelles, signalling pathways and the bioenergetics machinery of the infected cell to replicate their genome, produce viral proteins and assemble new viral particles. Mitochondria are key eukaryotic organelles often referred to as the cell's powerhouse. They control many fundamental cellular processes, from metabolism and energy production to calcium homeostasis and programmed cell death. Importantly, mitochondrial membranes are also critical sites for the integration and amplification of antiviral innate immune responses. Overall, mitochondria are therefore both supporting the virus life cycle by sustaining energy production, metabolism and synthesis of macromolecules and part of the cell's first line of defence against viruses. This review summarizes recent findings on viral manipulations of mitochondria and their functions. We explore the evolving understanding of how mitochondrial dynamics is targeted to regulate innate immunity, evasion strategies used to avoid mitochondrial-associated mechanisms that impair replication and the role of mitochondrial functions such as generating reactive oxygen species or regulating the electron transport chain during infection. Overall, we provide a comprehensive view of how viruses modulate mitochondrial function to promote replication.</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12282254/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144584121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Summary of taxonomy changes ratified by the International Committee on Taxonomy of Viruses (ICTV) from the Archaeal Viruses Subcommittee, 2025.","authors":"Mart Krupovic, Diana P Baquero, Eduardo A Bignon, Ariane Bize, Guillaume Borrel, Mingwei Cai, Lanming Chen, Marion Coves, Changhai Duan, Simonetta Gribaldo, Eugene V Koonin, Meng Li, Lirui Liu, Yang Liu, Sofia Medvedeva, Yimin Ni, Apoorva Prabhu, Christian Rinke, Yongjie Wang, Tianqi Xu, Shuling Yan, Qinglu Zeng, Rui Zhang, Ictv Taxonomy Summary Consortium","doi":"10.1099/jgv.0.002117","DOIUrl":"10.1099/jgv.0.002117","url":null,"abstract":"<p><p>The International Committee on Taxonomy of Viruses (ICTV) holds a ratification vote annually following the review of newly proposed taxa by ICTV Study Groups and members of the virology community. This article reports changes to the taxonomy of viruses infecting archaea that were approved and ratified by the ICTV in March 2025. Six new families of head-tailed viruses expanded the order <i>Caudoviricetes</i> (realm <i>Duplodnaviria</i>); one new family of filamentous viruses was added to the order <i>Ligamenvirales</i> (realm <i>Adnaviria</i>); one new family of viruses with pleomorphic virions was included within a new phylum, new order and new class in the kingdom <i>Trapavirae</i> (realm <i>Monodnaviria</i>); finally, three new families were created for spindle-shaped viruses that remain unassigned to higher level taxa. The 25 new species represent viruses infecting a broad range of archaea, including members of the classes Archaeoglobi, Bathyarchaeia, Methanobacteria, Methanomicrobia, Nitrososphaeria and Poseidoniia. Most of these viruses have been discovered by metagenomics in samples derived from diverse environments, including ambient and extreme marine ecosystems, the gastrointestinal tract of humans and animals, anaerobic digesters and terrestrial hot springs. Following this taxonomic update, archaeal viruses are officially classified into a total of 163 virus species in 94 genera within 62 families.</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144715087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Precision engineering of human cytomegalovirus without BAC constraints: a Sendai virus-delivered CRISPR/Cas9 approach.","authors":"Jillian C Carmichael, Christian S Stevens, Kristina E Atanasoff, Shreyas Kowdle, Rebecca A Reis, Domenico Tortorella, Benhur Lee","doi":"10.1099/jgv.0.002126","DOIUrl":"10.1099/jgv.0.002126","url":null,"abstract":"<p><p>Human cytomegalovirus (HCMV) genetic manipulation traditionally relies on bacterial artificial chromosome (BAC) recombineering, necessitated by its large ~236 kb genome. This approach is limited by the scarcity of HCMV strains engineered into BACs and often requires the deletion of 'non-essential' genes to accommodate the BAC cassette. We developed a novel approach using temperature-sensitive Sendai virus (SeV) vectors to deliver CRISPR/Cas9 for targeted HCMV genome editing without these constraints. This system achieves high editing efficiency (80-90%) in fibroblasts, epithelial cells and endothelial cells without BAC intermediates. As proof of principle, we targeted the HCMV (TB40/E strain) pentamer complex (PC) genes UL128 and UL130, crucial for viral entry into non-fibroblast cells. Edited viruses showed significantly reduced infectivity in epithelial cells, confirming functional disruption of the PC. Plaque purification yielded isogenic clones with phenotypes comparable to AD169, a naturally PC-deficient strain. Furthermore, multiplexed editing created precise 663 bp deletions in over 60% of viral genomes. Importantly, this method enables HCMV editing in physiologically relevant cell types without fibroblast passaging, which typically introduces mutations. This SeV-Cas9 system represents a significant advancement for studying HCMV biology in diverse cell types.</p>","PeriodicalId":15880,"journal":{"name":"Journal of General Virology","volume":"106 7","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12311846/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144642753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}