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Here we show that the spike protein of Omicron has a higher affinity for ACE2 compared with Delta, and a marked change in its antigenicity increases Omicron\u2019s evasion of therapeutic monoclonal and vaccine-elicited polyclonal neutralizing antibodies after two doses. mRNA vaccination as a third vaccine dose rescues and broadens neutralization. Importantly, the antiviral drugs remdesivir and molnupiravir retain efficacy against Omicron BA.1. Replication was similar for Omicron and Delta virus isolates in human nasal epithelial cultures. However,\u00a0in lung cells and gut cells, Omicron demonstrated lower replication. Omicron spike protein was less efficiently cleaved compared with Delta. The differences in replication were mapped to the entry efficiency of the virus on the basis\u00a0of spike-pseudotyped\u00a0virus assays. The defect in entry of Omicron pseudotyped virus to specific cell types effectively correlated with higher cellular RNA expression of\n                    <jats:italic>TMPRSS2<\/jats:italic>\n                    , and deletion of\n                    <jats:italic>TMPRSS2<\/jats:italic>\n                    affected Delta entry to a greater extent than Omicron. Furthermore, drug inhibitors targeting specific entry pathways\n                    <jats:sup>3<\/jats:sup>\n                    demonstrated that the Omicron spike inefficiently uses the cellular protease TMPRSS2, which promotes cell entry through plasma membrane fusion, with greater dependency on cell entry through the endocytic pathway. Consistent with suboptimal S1\/S2 cleavage and inability to use TMPRSS2, syncytium formation by the Omicron spike was substantially impaired compared with the Delta spike. The less efficient spike cleavage of Omicron at S1\/S2 is associated with a shift in cellular tropism away from TMPRSS2-expressing cells, with implications for altered pathogenesis.\n                  <\/jats:p>","DOI":"10.1038\/s41586-022-04474-x","type":"journal-article","created":{"date-parts":[[2022,2,1]],"date-time":"2022-02-01T11:59:30Z","timestamp":1643716770000},"page":"706-714","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1080,"title":["Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts infectivity and fusogenicity"],"prefix":"10.1038","volume":"603","author":[{"given":"Bo","family":"Meng","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam","family":"Abdullahi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabella A. T. 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