{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,28]],"date-time":"2026-02-28T04:31:29Z","timestamp":1772253089948,"version":"3.50.1"},"reference-count":62,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,8,22]],"date-time":"2024-08-22T00:00:00Z","timestamp":1724284800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000001","name":"NSF","doi-asserted-by":"publisher","award":["2026657"],"award-info":[{"award-number":["2026657"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"NSF","doi-asserted-by":"publisher","award":["2102948"],"award-info":[{"award-number":["2102948"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"NSF","doi-asserted-by":"publisher","award":["R56 AI150474-06A1"],"award-info":[{"award-number":["R56 AI150474-06A1"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"NSF","doi-asserted-by":"publisher","award":["\u0414O1-166"],"award-info":[{"award-number":["\u0414O1-166"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"name":"NIH","award":["2026657"],"award-info":[{"award-number":["2026657"]}]},{"name":"NIH","award":["2102948"],"award-info":[{"award-number":["2102948"]}]},{"name":"NIH","award":["R56 AI150474-06A1"],"award-info":[{"award-number":["R56 AI150474-06A1"]}]},{"name":"NIH","award":["\u0414O1-166"],"award-info":[{"award-number":["\u0414O1-166"]}]},{"name":"Ministry of Education and Science","award":["2026657"],"award-info":[{"award-number":["2026657"]}]},{"name":"Ministry of Education and Science","award":["2102948"],"award-info":[{"award-number":["2102948"]}]},{"name":"Ministry of Education and Science","award":["R56 AI150474-06A1"],"award-info":[{"award-number":["R56 AI150474-06A1"]}]},{"name":"Ministry of Education and Science","award":["\u0414O1-166"],"award-info":[{"award-number":["\u0414O1-166"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The efficiency of virus internalization into target cells is a major determinant of infectivity. SARS-CoV-2 internalization occurs via S-protein-mediated cell binding followed either by direct fusion with the plasma membrane or endocytosis and subsequent fusion with the endosomal membrane. Despite the crucial role of virus internalization, the precise kinetics of the processes involved remains elusive. We developed a pipeline, which combines live-cell microscopy and advanced image analysis, for measuring the rates of multiple internalization-associated molecular events of single SARS-CoV-2-virus-like particles (VLPs), including endosome ingression and pH change. Our live-cell imaging experiments demonstrate that only a few minutes after binding to the plasma membrane, VLPs ingress into RAP5-negative endosomes via dynamin-dependent scission. Less than two minutes later, VLP speed increases in parallel with a pH drop below 5, yet these two events are not interrelated. By co-imaging fluorescently labeled nucleocapsid proteins, we show that nucleocapsid release occurs with similar kinetics to VLP acidification. Neither Omicron mutations nor abrogation of the S protein polybasic cleavage site affected the rate of VLP internalization, indicating that they do not confer any significant advantages or disadvantages during this process. Finally, we observe that VLP internalization occurs two to three times faster in VeroE6 than in A549 cells, which may contribute to the greater susceptibility of the former cell line to SARS-CoV-2 infection. Taken together, our precise measurements of the kinetics of VLP internalization-associated processes shed light on their contribution to the effectiveness of SARS-CoV-2 propagation in cells.<\/jats:p>","DOI":"10.3390\/v16081341","type":"journal-article","created":{"date-parts":[[2024,8,22]],"date-time":"2024-08-22T03:07:38Z","timestamp":1724296058000},"page":"1341","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Kinetic Landscape of Single Virus-like Particles Highlights the Efficacy of SARS-CoV-2 Internalization"],"prefix":"10.3390","volume":"16","author":[{"given":"Aleksandar","family":"Atemin","sequence":"first","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 21, G. Bontchev Str., 1113 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aneliya","family":"Ivanova","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 21, G. Bontchev Str., 1113 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wiley","family":"Peppel","sequence":"additional","affiliation":[{"name":"Department of Physics and Astronomy, University of Utah, Salt Lake City, UT 84112, USA"},{"name":"Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rumen","family":"Stamatov","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 21, G. Bontchev Str., 1113 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rodrigo","family":"Gallegos","sequence":"additional","affiliation":[{"name":"Department of Physics and Astronomy, University of Utah, Salt Lake City, UT 84112, USA"},{"name":"Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haley","family":"Durden","sequence":"additional","affiliation":[{"name":"Department of Physics and Astronomy, University of Utah, Salt Lake City, UT 84112, USA"},{"name":"Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sonya","family":"Uzunova","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 21, G. Bontchev Str., 1113 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael D.","family":"Vershinin","sequence":"additional","affiliation":[{"name":"Department of Physics and Astronomy, University of Utah, Salt Lake City, UT 84112, USA"},{"name":"Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA"},{"name":"Department of Biology, University of Utah, Salt Lake City, UT 84112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9186-1230","authenticated-orcid":false,"given":"Saveez","family":"Saffarian","sequence":"additional","affiliation":[{"name":"Department of Physics and Astronomy, University of Utah, Salt Lake City, UT 84112, USA"},{"name":"Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA"},{"name":"Department of Biology, University of Utah, Salt Lake City, UT 84112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stoyno S.","family":"Stoynov","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 21, G. Bontchev Str., 1113 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"106054","DOI":"10.1016\/j.ijantimicag.2020.106054","article-title":"Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2): A global pandemic and treatment strategies","volume":"56","author":"Sharma","year":"2020","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"138882","DOI":"10.1016\/j.scitotenv.2020.138882","article-title":"COVID-19 outbreak: Migration, effects on society, global environment and prevention","volume":"728","author":"Chakraborty","year":"2020","journal-title":"Sci. Total Environ."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1016\/S0140-6736(20)30557-2","article-title":"SARS-CoV-2 is an appropriate name for the new coronavirus","volume":"395","author":"Wu","year":"2020","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Coronaviridae Study Group of the International Committee on Taxonomy of Viruses (2020). The species Severe acute respiratory syndrome-related coronavirus: Classifying 2019-nCoV and naming it SARS-CoV-2. Nat. Microbiol., 5, 536\u2013544.","DOI":"10.1038\/s41564-020-0695-z"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/j.chom.2020.02.001","article-title":"Genome Composition and Divergence of the Novel Coronavirus (2019-nCoV) Originating in China","volume":"27","author":"Wu","year":"2020","journal-title":"Cell Host Microbe"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Siddell, S.G. (1995). Coronavirus Gene Expression. The Coronaviridae, Springer US.","DOI":"10.1007\/978-1-4899-1531-3"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1016\/j.arcmed.2020.05.012","article-title":"Structural Proteins in Severe Acute Respiratory Syndrome Coronavirus-2","volume":"51","author":"Satarker","year":"2020","journal-title":"Arch. Med. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1626","DOI":"10.1126\/science.abl6184","article-title":"Rapid assessment of SARS-CoV-2\u2013evolved variants using virus-like particles","volume":"374","author":"Syed","year":"2021","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Nooraei, S., Bahrulolum, H., Hoseini, Z.S., Katalani, C., Hajizade, A., Easton, A.J., and Ahmadian, G. (2021). Virus-like particles: Preparation, immunogenicity and their roles as nanovaccines and drug nanocarriers. J. Nanobiotechnol., 19.","DOI":"10.1186\/s12951-021-00806-7"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Xu, R., Shi, M., Li, J., Song, P., and Li, N. (2020). Construction of SARS-CoV-2 Virus-Like Particles by Mammalian Expression System. Front. Bioeng. Biotechnol., 8.","DOI":"10.3389\/fbioe.2020.00862"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Swann, H., Sharma, A., Preece, B., Peterson, A., Eldridge, C., Belnap, D.M., Vershinin, M., and Saffarian, S. (2020). Minimal system for assembly of SARS-CoV-2 virus like particles. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-78656-w"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Gourdelier, M., Swain, J., Arone, C., Mouttou, A., Bracquemond, D., Merida, P., Saffarian, S., Lyonnais, S., Favard, C., and Muriaux, D. (2022). Optimized production and fluorescent labeling of SARS-CoV-2 virus-like particles. Sci. Rep., 12.","DOI":"10.1038\/s41598-022-18681-z"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.bbrc.2020.11.080","article-title":"Structural stability of SARS-CoV-2 virus like particles degrades with temperature","volume":"534","author":"Sharma","year":"2021","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1038\/s41580-021-00418-x","article-title":"Mechanisms of SARS-CoV-2 entry into cells","volume":"23","author":"Jackson","year":"2022","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"872","DOI":"10.1053\/j.gastro.2008.10.055","article-title":"Tissue-specific amino acid transporter partners ACE2 and collectrin differentially interact with hartnup mutations","volume":"136","author":"Camargo","year":"2009","journal-title":"Gastroenterology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1016\/j.cell.2020.09.033","article-title":"SARS-CoV-2 Infection Depends on Cellular Heparan Sulfate and ACE2","volume":"183","author":"Clausen","year":"2020","journal-title":"Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1038\/s42255-020-00324-0","article-title":"HDL-scavenger receptor B type 1 facilitates SARS-CoV-2 entry","volume":"2","author":"Wei","year":"2020","journal-title":"Nat. Metab."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"856","DOI":"10.1126\/science.abd2985","article-title":"Neuropilin-1 facilitates SARS-CoV-2 cell entry and infectivity","volume":"370","author":"Ojha","year":"2020","journal-title":"Science"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1126\/science.abd3072","article-title":"Neuropilin-1 is a host factor for SARS-CoV-2 infection","volume":"370","author":"Daly","year":"2020","journal-title":"Science"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1086\/427811","article-title":"Function of HAb18G\/CD147 in invasion of host cells by severe acute respiratory syndrome coronavirus","volume":"191","author":"Chen","year":"2005","journal-title":"J. Infect. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1038\/s41392-020-00426-x","article-title":"CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells","volume":"5","author":"Wang","year":"2020","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1038\/s41422-020-00460-y","article-title":"AXL is a candidate receptor for SARS-CoV-2 that promotes infection of pulmonary and bronchial epithelial cells","volume":"31","author":"Wang","year":"2021","journal-title":"Cell Res."},{"key":"ref_23","first-page":"2020-09","article-title":"KIM-1\/TIM-1 is a Receptor for SARS-CoV-2 in Lung and Kidney SARS-CoV-2 precipitates respiratory distress by infection of airway epithelial cells and is often domain 1 (KIM-1\/TIM-1) is expressed in lung and kidney epithelial cells in COVID-19 patient","volume":"2","author":"Mori","year":"2022","journal-title":"MedRxiv"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"15748","DOI":"10.1073\/pnas.0403812101","article-title":"CD209L (L-SIGN) is a receptor for severe acute respiratory syndrome coronavirus","volume":"101","author":"Jeffers","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1156","DOI":"10.1021\/acscentsci.0c01537","article-title":"CD209L L-SIGN and CD209 DC-SIGN act as receptors for SARS-CoV-2 amraei","volume":"7","author":"Amraei","year":"2021","journal-title":"ACS Cent. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6997","DOI":"10.1039\/D2SC06967A","article-title":"The ACE2 receptor accelerates but is not biochemically required for SARS-CoV-2 membrane fusion","volume":"14","author":"Cervantes","year":"2023","journal-title":"Chem. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1038\/s41579-020-00468-6","article-title":"Coronavirus biology and replication: Implications for SARS-CoV-2","volume":"19","author":"Kratzel","year":"2021","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Bayati, A., Kumar, R., Francis, V., and McPherson, P.S. (2021). SARS-CoV-2 infects cells after viral entry via clathrin-mediated endocytosis. J. Biol. Chem., 296.","DOI":"10.1016\/j.jbc.2021.100306"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1021\/bi00052a046","article-title":"Kinetics of the pH-induced inactivation of human cathepsin L","volume":"32","author":"Turk","year":"1993","journal-title":"Biochemistry"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1038\/s41392-021-00558-8","article-title":"Cathepsin L plays a key role in SARS-CoV-2 infection in humans and humanized mice and is a promising target for new drug development","volume":"6","author":"Zhao","year":"2021","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"eadd5446","DOI":"10.1126\/sciimmunol.add5446","article-title":"An antibody from single human VH-rearranging mouse neutralizes all SARS-CoV-2 variants through BA.5 by inhibiting membrane fusion","volume":"7","author":"Luo","year":"2022","journal-title":"Sci. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e2209514119","DOI":"10.1073\/pnas.2209514119","article-title":"SARS-CoV-2 requires acidic pH to infect cells","volume":"119","author":"Kreutzberger","year":"2022","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2199","DOI":"10.1016\/S0006-3495(00)76468-X","article-title":"The use of pHluorins for optical measurements of presynaptic activity","volume":"79","author":"Sankaranarayanan","year":"2000","journal-title":"Biophys. J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2210","DOI":"10.1016\/j.scib.2023.08.031","article-title":"Single particle tracking reveals SARS-CoV-2 regulating and utilizing dynamic filopodia for viral invasion","volume":"68","author":"Zhang","year":"2023","journal-title":"Sci. Bull."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s13567-017-0412-y","article-title":"Betanodavirus-like particles enter host cells via clathrin-mediated endocytosis in a cholesterol-, pH- and cytoskeleton-dependent manner","volume":"48","author":"Huang","year":"2017","journal-title":"Vet. Res."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Zepeda-Cervantes, J., Ram\u00edrez-Jarqu\u00edn, J.O., and Vaca, L. (2020). Interaction Between Virus-Like Particles (VLPs) and Pattern Recognition Receptors (PRRs) From Dendritic Cells (DCs): Toward Better Engineering of VLPs. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.01100"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1099\/jgv.0.001453","article-title":"SARS-coronavirus-2 replication in Vero E6 cells: Replication kinetics, rapid adaptation and cytopathology","volume":"101","author":"Ogando","year":"2020","journal-title":"J. Gen. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Prichard, K.L., O\u2019Brien, N.S., Murcia, S.R., Baker, J.R., and McCluskey, A. (2022). Role of Clathrin and Dynamin in Clathrin Mediated Endocytosis\/Synaptic Vesicle Recycling and Implications in Neurological Diseases. Front. Cell. Neurosci., 15.","DOI":"10.3389\/fncel.2021.754110"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3595","DOI":"10.1091\/mbc.e14-07-1240","article-title":"Dynamin recruitment and membrane scission at the neck of a clathrin-coated pit","volume":"25","author":"Cocucci","year":"2014","journal-title":"Mol. Biol. Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2270","DOI":"10.15252\/embj.201694613","article-title":"Membrane fission by dynamin: What we know and what we need to know","volume":"35","author":"Antonny","year":"2016","journal-title":"EMBO J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1038\/s41556-021-00713-x","article-title":"Dynamin-dependent vesicle twist at the final stage of clathrin-mediated endocytosis","volume":"23","author":"Cheng","year":"2021","journal-title":"Nat. Cell Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1083\/jcb.152.2.309","article-title":"The role of dynamin and its binding partners in coated pit invagination and scission","volume":"152","author":"Hill","year":"2001","journal-title":"J. Cell Biol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"\u0160timac, I., Vu\u010dko, N.J., Zagorac, G.B., Marceli\u0107, M., Lu\u010din, H.M., and Lu\u010din, P. (2021). Dynamin inhibitors prevent the establishment of the cytomegalovirus assembly compartment in the early phase of infection. Life, 11.","DOI":"10.3390\/life11090876"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"0628","DOI":"10.1371\/journal.pbio.1000057","article-title":"Cargo and dynamin regulate clathrin-coated pit maturation","volume":"7","author":"Loerke","year":"2009","journal-title":"PLoS Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.addr.2016.08.014","article-title":"Investigation of endosome and lysosome biology by ultra pH-sensitive nanoprobes","volume":"113","author":"Wang","year":"2017","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.chroma.2015.07.019","article-title":"Adsorption of virus-like particles on ion exchange surface: Conformational changes at different pH detected by dual polarization interferometry","volume":"1408","author":"Yang","year":"2015","journal-title":"J. Chromatogr. A"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"19478","DOI":"10.1074\/jbc.M603313200","article-title":"conformational stability and disassembly of norwalk virus-like particles: Effect of ph and temperature","volume":"281","author":"Ausar","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e1802081","DOI":"10.1002\/smll.201802081","article-title":"Experimental and Theoretical Determination of the pH inside the Confinement of a Virus-Like Particle","volume":"14","author":"Maassen","year":"2018","journal-title":"Small"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1412","DOI":"10.1038\/s41467-017-01752-5","article-title":"Semisynthetic fluorescent pH sensors for imaging exocytosis and endocytosis","volume":"8","author":"Martineau","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1099\/0022-1317-43-1-247","article-title":"Regulation of the interferon system: Evidence that Vero cells have a genetic defect in interferon production","volume":"43","author":"Emeny","year":"1979","journal-title":"J. Gen. Virol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1380","DOI":"10.3201\/eid2705.210023","article-title":"Susceptibility to SARS-CoV-2 of cell lines and substrates commonly used to diagnose and isolate influenza and other viruses","volume":"27","author":"Wang","year":"2021","journal-title":"Emerg. Infect. Dis."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"e2","DOI":"10.1016\/S2666-5247(20)30008-2","article-title":"SARS-CoV-2 cellular tropism","volume":"1","author":"Cagno","year":"2020","journal-title":"Lancet Microbe"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1007\/s15010-020-01486-5","article-title":"Differences and similarities between SARS-CoV and SARS-CoV-2: Spike receptor-binding domain recognition and host cell infection with support of cellular serine proteases","volume":"48","author":"Rossi","year":"2020","journal-title":"Infection"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1080\/22221751.2020.1756700","article-title":"Attenuated SARS-CoV-2 variants with deletions at the S1\/S2 junction","volume":"9","author":"Lau","year":"2020","journal-title":"Emerg. Microbes Infect."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Sasaki, M., Uemura, K., Sato, A., Toba, S., Sanaki, T., Maenaka, K., Hall, W.W., Orba, Y., and Sawa, H. (2021). SARS-CoV-2 variants with mutations at the S1\/ S2 cleavage site are generated in vitro during propagation in TMPRSS2-deficient cells. PLoS Pathog., 17.","DOI":"10.1101\/2020.08.28.271163"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1038\/s41579-021-00573-0","article-title":"SARS-CoV-2 variants, spike mutations and immune escape","volume":"19","author":"Harvey","year":"2021","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"899","DOI":"10.1038\/s41564-021-00908-w","article-title":"The furin cleavage site in the SARS-CoV-2 spike protein is required for transmission in ferrets","volume":"6","author":"Peacock","year":"2021","journal-title":"Nat. Microbiol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"e570","DOI":"10.1016\/S2666-5247(23)00144-1","article-title":"The SARS-CoV-2 furin cleavage site: Natural selection or smoking gun?","volume":"4","author":"Lubinski","year":"2023","journal-title":"Lancet Microbe"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1038\/s41586-022-04479-6","article-title":"SARS-CoV-2 Omicron variant replication in human bronchus and lung ex vivo","volume":"603","author":"Hui","year":"2022","journal-title":"Nature"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Letoha, A., Hud\u00e1k, A., and Letoha, T. (2023). Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241814140"},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Hud\u00e1k, A., Letoha, A., Szil\u00e1k, L., and Letoha, T. (2021). Contribution of syndecans to the cellular entry of SARS-CoV-2. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22105336"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1016\/j.molcel.2020.04.022","article-title":"A Multibasic Cleavage Site in the Spike Protein of SARS-CoV-2 Is Essential for Infection of Human Lung Cells","volume":"78","author":"Hoffmann","year":"2020","journal-title":"Mol. Cell"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/16\/8\/1341\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:40:48Z","timestamp":1760110848000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/16\/8\/1341"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,22]]},"references-count":62,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2024,8]]}},"alternative-id":["v16081341"],"URL":"https:\/\/doi.org\/10.3390\/v16081341","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2024.06.10.598174","asserted-by":"object"}]},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,8,22]]}}}