{"title":"Cryo-electron tomography of viral infection — from applications to biosafety","authors":"Liv Zimmermann, Petr Chlanda","doi":"10.1016/j.coviro.2023.101338","DOIUrl":null,"url":null,"abstract":"<div><p>Cellular cryo-electron tomography (cryo-ET) offers 3D snapshots at molecular resolution capturing pivotal steps during viral infection. However, tomogram quality depends on the vitrification level of the sample and its thickness. In addition, mandatory inactivation protocols to assure biosafety when handling highly pathogenic viruses during cryo-ET can compromise sample preservation. Here, we focus on different strategies applied in cryo-ET and discuss their advantages and limitations with reference to severe acute respiratory syndrome coronavirus 2 studies. We highlight the importance of virus-like particle (VLP) and replicon systems to study virus assembly and replication in a cellular context without inactivation protocols. We discuss the application of chemical fixation and different irradiation methods in cryo-ET sample preparation and acquisition workflows.</p></div>","PeriodicalId":11082,"journal":{"name":"Current opinion in virology","volume":"61 ","pages":"Article 101338"},"PeriodicalIF":5.7000,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current opinion in virology","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S187962572300038X","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"VIROLOGY","Score":null,"Total":0}
引用次数: 1
Abstract
Cellular cryo-electron tomography (cryo-ET) offers 3D snapshots at molecular resolution capturing pivotal steps during viral infection. However, tomogram quality depends on the vitrification level of the sample and its thickness. In addition, mandatory inactivation protocols to assure biosafety when handling highly pathogenic viruses during cryo-ET can compromise sample preservation. Here, we focus on different strategies applied in cryo-ET and discuss their advantages and limitations with reference to severe acute respiratory syndrome coronavirus 2 studies. We highlight the importance of virus-like particle (VLP) and replicon systems to study virus assembly and replication in a cellular context without inactivation protocols. We discuss the application of chemical fixation and different irradiation methods in cryo-ET sample preparation and acquisition workflows.
期刊介绍：
Current Opinion in Virology (COVIRO) is a systematic review journal that aims to provide specialists with a unique and educational platform to keep up to date with the expanding volume of information published in the field of virology. It publishes 6 issues per year covering the following 11 sections, each of which is reviewed once a year: Emerging viruses: interspecies transmission; Viral immunology; Viral pathogenesis; Preventive and therapeutic vaccines; Antiviral strategies; Virus structure and expression; Animal models for viral diseases; Engineering for viral resistance; Viruses and cancer; Virus vector interactions. There is also a section that changes every year to reflect hot topics in the field.
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