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This mutation is in linkage disequilibrium with an ORF1b protein variant (P314L), making it difficult to discern the functional significance of the Spike D614G mutation from population genetics alone. Here, we perform site-directed mutagenesis on wild-type human-codon-optimized Spike to introduce the D614G variant. Using multiple human cell lines, including human lung epithelial cells, we found that the lentiviral particles pseudotyped with Spike D614G are more effective at transducing cells than ones pseudotyped with wild-type Spike. The increased transduction with Spike D614G ranged from 1.3- to 2.4-fold in Caco-2 and Calu-3 cells expressing endogenous ACE2 and from 1.5- to 7.7-fold in A549\n                    <jats:sup>ACE2<\/jats:sup>\n                    and Huh7.5\n                    <jats:sup>ACE2<\/jats:sup>\n                    overexpressing ACE2. Furthermore,\n                    <jats:italic>trans<\/jats:italic>\n                    -complementation of SARS-CoV-2 virus with Spike D614G showed an increased infectivity in human cells. Although there is minimal difference in ACE2 receptor binding between the D614 and G614 Spike variants, the G614 variant is more resistant to proteolytic cleavage, suggesting a possible mechanism for the increased transduction.\n                  <\/jats:p>","DOI":"10.7554\/elife.65365","type":"journal-article","created":{"date-parts":[[2021,2,11]],"date-time":"2021-02-11T08:02:32Z","timestamp":1613030552000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.65365","source":"Crossref","is-referenced-by-count":166,"title":["The Spike D614G mutation increases SARS-CoV-2 infection of multiple human cell types"],"prefix":"10.7554","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3453-0849","authenticated-orcid":true,"given":"Zharko","family":"Daniloski","sequence":"first","affiliation":[{"name":"New York Genome Center, New York, United States"},{"name":"Department of Biology, New York University, New York, United 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