Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts infectivity and fusogenicity methods
Show snapshot details
On this page
This experiment, in seven questions
Jump straight to the part of the recipe you need. Data and provenance labels stay close to the action they support.
Shopping and prep list
What do I need before I start?
Biological model pending
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Before you run
What should be confirmed before execution?
First confirmation
Species or subject information is missing.
Confirm before execution
Protocol does not list structured equipment.
Confirm before execution
This page shows passages quoted from the paper. Equipment, materials, times, and temperatures are hidden until the page is rebuilt from the full paper and checked by a reviewer. Check the paper before using this method.
Confirm before execution
Open the source paper before finalizing run-specific details.
Procurement checkpoint
Use source-stated vendors where present. Treat mapped products as sourcing options unless the page marks an exact source match.
Open quote workflowStep-by-step procedure
What do I do, in order?
The sensitivity of Omicron to therapeutics
Omicron was predicted to have broad resistance to neutralizing antibodies based on mutations in the class I-IV antigenic regions in its RBD. Current standard of care antiviral treatment for moderate to severe COVID-19 includes the use of the monoclonal antibody combination REGN10933 (casivirimab) and REGN10897 (imdevimab). The K417N, E484A, S477N and Q493R mutations preclude REGN10933 binding, whereas G446S clashes with REGN10987, consistent with the decreased binding to the Omicron RBD and S trimer and the loss of neutralization previously descibed,. We next tested serial dilutions of component monoclonal antibodies, both individually and in combination, against Delta and Omicron live viruses in tissue culture (Fig. ). Although the Delta variant was effectively neutralized by casirivimab, imdevimab was only partially effective, consistent with previous data. In co...
Omicron and polyclonal antibodies
An important question is whether vaccine-elicited antibodies are able to neutralize Omicron. We synthesized codon-optimized spike expression plasmids for Omicron and Delta spike proteins and generated pseudotyped virus (PV) particles by co-transfecting the spike expression plasmids with a lentiviral gag-pol-expressing plasmid and a lentiviral transduction plasmid encoding the luciferase gene,. We obtained longitudinal serum samples from 40 individuals who were vaccinated with either the BNT162b2 or ChAdOx-1 vaccine and performed serum titrations before mixing sera with our reporter PV particles. The participants had a median age of around 70 years and prospective serum samples were taken as follows: one month after dose two, six months after dose two and one month after dose three (Extended Data Table ). We observed at least tenfold loss of neutralization against Omicron after the s...
Omicron and polyclonal antibodies
To confirm the loss of neutralizing activity against Omicron after the second dose, we next used a live virus experimental system to compare Delta and Omicron variants against sera taken four weeks after the second dose of BNT162b2, and obtained similar results to those obtained in the PV assay (Extended Data Fig. ). These live viruses were also used to assess the neutralization of the Omicron variant by sera derived from unvaccinated individuals who were previously infected with the early Wuhan-Hu-1 virus or the Delta variant. As expected, vaccine sera had significantly impaired activity against Omicron compared with Delta (Extended Data Fig. ). We also tested mRNA-1273 vaccine-elicited sera, which showed a similar reduction in neutralization to BNT162b2. However, coronavac sera showed little neutralization against Delta and 0 out of 9 participants had detectable neutralization again...
Omicron replication and spike cleavage
We infected primary human nasal epithelial 3D cultures (hNECs) with an air-liquid interface (Fig. ). Infection with live SARS-CoV-2 Omicron or Delta was conducted at the apical surface with equal amounts of input virus and virus collected from the apical surface at 24 and 48 h, and virus was quantified using quantitative PCR (qPCR) analysis of the E gene and 50% tissue culture infectious dose (TCID 50 ). We observed similar replication kinetics for Omicron and Delta in the hNECs (Fig. ). We next infected Calu-3 lung cells (which are known to express endogenous TMPRSS2 ) with live isolates, and observed significantly greater viral replication for Delta than Omicron (Fig. ), manifesting as early as 24 h after infection. We also infected Caco-2 cells, a colon cancer cell line (known to express high levels of endogenous TMPRSS2 ) and, as for Calu-3, we found 1 log...
Impaired cell-cell fusion by Omicron S
The ability of viral membrane glycoproteins to induce cell-cell fusion and syncytium formation is well established -, providing an additional route for SARS-CoV-2 dissemination that may facilitate evasion of neutralizing antibodies. The role of syncytium formation in viral replication and the pathogenesis of severe COVID-19 has been reported and may be a druggable process to treat COVID-19 pathology,. SARS-CoV-2-mediated cell-cell fusion requires the PBCS and spike cleavage at S1/S2, and the process is known to be accelerated by the presence of TMPRSS2 (ref. ).
Impaired cell-cell fusion by Omicron S
Given the requirement of TMPRSS2 for optimal cell-cell fusion, we hypothesized that Omicron may be impaired in mediating this process. We used a split GFP system to monitor cell-cell fusion in real time (Fig. ). As a control to show that spike cleavage is needed for fusion in our assay system, we titrated CMK (a furin inhibitor) into donor cell medium before transfection with spike-expressing plasmids, observing dose-dependent inhibition of fusion (Extended Data Fig. ). As a further control to demonstrate the need for ACE2 engagement by expressed spike, we tested the ability of convalescent serum containing neutralizing antibodies to block cell-cell fusion. Indeed, serum blocked syncytia formation in a dose-dependent manner (Extended Data Fig. ). Fig. 4 SARS-CoV-2 Omicron variant spike shows impaired cell-cell fusion activity and smaller infection foci generate...
Impaired cell-cell fusion by Omicron S
We predicted that poor fusion would impair cell-cell spread of Omicron, and analysed infection focus size using live virus infection of H1299 cells, in which we have shown similar entry efficiency for Omicron and Delta spike PV (Fig. ). Infection foci in spreading virus infections occur because of localized (cell-to-cell) infection, probably facilitated by syncytium formation. The infection was followed by overlay of semi-solid medium (carboxymethylcellulose) to inhibit cell-free infection, thereby favouring cell-cell infection. Infected cells were stained with an anti-S antibodies, and local spread was characterized by the staining of foci visible to the naked eye (Fig. ). Omicron infection resulted in slightly higher numbers of foci (Fig. ), but each infection focus was substantially smaller in size relative to those formed by Delta infection (Fig. ). For a more detailed...
Flow cytometry analysis of spike protein surface expression
ExpiCHO cells were seeded into 50 ml Mini Bioreactor Centrifuge Tube (Corning) at 6 × 10 6 cells per ml in 5 ml ExpiCHO Expression medium (Life technology). Plasmids encoding SARS-CoV-2 Wuhan, Alpha (B.1.1.7), Delta (B.1.617.2) or Omicron (B.1.1.529) spike protein (5 mg) were diluted in OptiPRO SFM (Life Technology) and mixed with Expifectamine CHO Reagent (Life Technology). After incubation for 1 min at room temperature, transfection mixes were added to the cell suspensions. Next, the cells were incubated at 37 °C under 8% CO 2 with an orbital shaking speed of 120 rpm for the next 48 h.
Measurement outputs
What raw and processed outputs should exist?
Analysis plan
How should the outputs become interpretable results?
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
Review raw outputs for quality, remove unusable captures, and organize the data into a comparison-ready table or image set.
inferred from protocolScoring or quantification
Quantify the primary endpoint described in the methods before moving into any statistical comparison.
inferred from protocolStatistical comparison
Statistical method not yet structured for this page.
inferred from protocolReporting output
Report representative outputs alongside the summary comparison used to support the conclusion.
inferred from protocolSource and audit
What supports the facts on this page?
Evidence quotes (8)
Omicron was predicted to have broad resistance to neutralizing antibodies based on mutations in the class I-IV antigenic regions in its RBD. Current standard of care antiviral treatment for moderate to severe COVID-19 includes the use of the monoclonal antibody combination REGN10933 (casivirimab) and REGN10897 (imdevimab). The K417N, E484A, S477N and Q493R mutations preclude REGN10933 binding, whereas G446S clashes with REGN10987, consistent with the decreased binding to the Omicron RBD and S trimer and the loss of neutralization previously descibed,. We next tested serial dilutions of component monoclonal antibodies, both individually and in combination, against Delta and Omicron live viruses in tissue culture (Fig. ). Although the Delta variant was effectively neutralized by casirivimab, imdevimab was only partially effective, consistent with previous data. In combination, these monoclonal antibodies were highly potent against Delta. However, there was a complete loss of neutralizing activity against Omicron by these monoclonal antibodies alone or in combination (Fig. ). Given these results, we next tested the direct-acting antivirals remdesivir and the act...
An important question is whether vaccine-elicited antibodies are able to neutralize Omicron. We synthesized codon-optimized spike expression plasmids for Omicron and Delta spike proteins and generated pseudotyped virus (PV) particles by co-transfecting the spike expression plasmids with a lentiviral gag-pol-expressing plasmid and a lentiviral transduction plasmid encoding the luciferase gene,. We obtained longitudinal serum samples from 40 individuals who were vaccinated with either the BNT162b2 or ChAdOx-1 vaccine and performed serum titrations before mixing sera with our reporter PV particles. The participants had a median age of around 70 years and prospective serum samples were taken as follows: one month after dose two, six months after dose two and one month after dose three (Extended Data Table ). We observed at least tenfold loss of neutralization against Omicron after the second dose compared with Delta (Fig. and Extended Data Fig. ). Indeed, neutralization of Omicron was not detectable for the majority of individuals who had received two doses of ChAdOx-1. We also observed waning over time after the second dose for both vaccines (Fig. ). Both groups received a booste...
To confirm the loss of neutralizing activity against Omicron after the second dose, we next used a live virus experimental system to compare Delta and Omicron variants against sera taken four weeks after the second dose of BNT162b2, and obtained similar results to those obtained in the PV assay (Extended Data Fig. ). These live viruses were also used to assess the neutralization of the Omicron variant by sera derived from unvaccinated individuals who were previously infected with the early Wuhan-Hu-1 virus or the Delta variant. As expected, vaccine sera had significantly impaired activity against Omicron compared with Delta (Extended Data Fig. ). We also tested mRNA-1273 vaccine-elicited sera, which showed a similar reduction in neutralization to BNT162b2. However, coronavac sera showed little neutralization against Delta and 0 out of 9 participants had detectable neutralization against Omicron. Sera from Delta infections appeared to have lower cross-neutralization compared with those from the early pandemic period when Wuhan-Hu-1 D614G was dominant (Extended Data Fig. ).
We infected primary human nasal epithelial 3D cultures (hNECs) with an air-liquid interface (Fig. ). Infection with live SARS-CoV-2 Omicron or Delta was conducted at the apical surface with equal amounts of input virus and virus collected from the apical surface at 24 and 48 h, and virus was quantified using quantitative PCR (qPCR) analysis of the E gene and 50% tissue culture infectious dose (TCID 50 ). We observed similar replication kinetics for Omicron and Delta in the hNECs (Fig. ). We next infected Calu-3 lung cells (which are known to express endogenous TMPRSS2 ) with live isolates, and observed significantly greater viral replication for Delta than Omicron (Fig. ), manifesting as early as 24 h after infection. We also infected Caco-2 cells, a colon cancer cell line (known to express high levels of endogenous TMPRSS2 ) and, as for Calu-3, we found 1 log greater Delta RNA and higher infectious virus (TCID 50 ) in the supernatants compared with those observed for Omicron. Finally, we tested a cell line-HeLa, overexpressing ACE2 and TMPRSS2 -and obtained similar results to those of the Calu-3 and Caco-2 cells. Fig. 2 SARS-CoV-2 Omicron and...
The ability of viral membrane glycoproteins to induce cell-cell fusion and syncytium formation is well established -, providing an additional route for SARS-CoV-2 dissemination that may facilitate evasion of neutralizing antibodies. The role of syncytium formation in viral replication and the pathogenesis of severe COVID-19 has been reported and may be a druggable process to treat COVID-19 pathology,. SARS-CoV-2-mediated cell-cell fusion requires the PBCS and spike cleavage at S1/S2, and the process is known to be accelerated by the presence of TMPRSS2 (ref. ).
Given the requirement of TMPRSS2 for optimal cell-cell fusion, we hypothesized that Omicron may be impaired in mediating this process. We used a split GFP system to monitor cell-cell fusion in real time (Fig. ). As a control to show that spike cleavage is needed for fusion in our assay system, we titrated CMK (a furin inhibitor) into donor cell medium before transfection with spike-expressing plasmids, observing dose-dependent inhibition of fusion (Extended Data Fig. ). As a further control to demonstrate the need for ACE2 engagement by expressed spike, we tested the ability of convalescent serum containing neutralizing antibodies to block cell-cell fusion. Indeed, serum blocked syncytia formation in a dose-dependent manner (Extended Data Fig. ). Fig. 4 SARS-CoV-2 Omicron variant spike shows impaired cell-cell fusion activity and smaller infection foci generated by live virus. a, Schematic of the cell-cell fusion assay. The schematic was created using BioRender.com. b, Spike expression at the cell surface as determined by flow cytometry, showing the distribution of fluorescence intensity. The percentage of spike-positive cells is indicated....
We predicted that poor fusion would impair cell-cell spread of Omicron, and analysed infection focus size using live virus infection of H1299 cells, in which we have shown similar entry efficiency for Omicron and Delta spike PV (Fig. ). Infection foci in spreading virus infections occur because of localized (cell-to-cell) infection, probably facilitated by syncytium formation. The infection was followed by overlay of semi-solid medium (carboxymethylcellulose) to inhibit cell-free infection, thereby favouring cell-cell infection. Infected cells were stained with an anti-S antibodies, and local spread was characterized by the staining of foci visible to the naked eye (Fig. ). Omicron infection resulted in slightly higher numbers of foci (Fig. ), but each infection focus was substantially smaller in size relative to those formed by Delta infection (Fig. ). For a more detailed view of the foci, we imaged at a higher magnification using a microscope objective, and used cells that were imaged 2 h after infection with ancestral virus-when relatively little viral transmission is expected to be completed-as an approximate measure of the size of a single cell...
ExpiCHO cells were seeded into 50 ml Mini Bioreactor Centrifuge Tube (Corning) at 6 × 10 6 cells per ml in 5 ml ExpiCHO Expression medium (Life technology). Plasmids encoding SARS-CoV-2 Wuhan, Alpha (B.1.1.7), Delta (B.1.617.2) or Omicron (B.1.1.529) spike protein (5 mg) were diluted in OptiPRO SFM (Life Technology) and mixed with Expifectamine CHO Reagent (Life Technology). After incubation for 1 min at room temperature, transfection mixes were added to the cell suspensions. Next, the cells were incubated at 37 °C under 8% CO 2 with an orbital shaking speed of 120 rpm for the next 48 h.
Machine-readable layer
[
{
"@context": "https://schema.org",
"@type": "HowTo",
"name": "Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts infectivity and fusogenicity methods",
"description": "Protocol for Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts infectivity and fusogenicity methods",
"step": [
{
"@type": "HowToStep",
"position": 1,
"name": "The sensitivity of Omicron to therapeutics",
"text": "Omicron was predicted to have broad resistance to neutralizing antibodies based on mutations in the class I-IV antigenic regions in its RBD. Current standard of care antiviral treatment for moderate to severe COVID-19 includes the use of the monoclonal antibody combination REGN10933 (casivirimab) and REGN10897 (imdevimab). The K417N, E484A, S477N and Q493R mutations preclude REGN10933 binding, whereas G446S clashes with REGN10987, consistent with the decreased binding to the Omicron RBD and S trimer and the loss of neutralization previously descibed,. We next tested serial dilutions of component monoclonal antibodies, both individually and in combination, against Delta and Omicron live viruses in tissue culture (Fig. ). Although the Delta variant was effectively neutralized by casirivimab, imdevimab was only partially effective, consistent with previous data. In co..."
},
{
"@type": "HowToStep",
"position": 2,
"name": "Omicron and polyclonal antibodies",
"text": "An important question is whether vaccine-elicited antibodies are able to neutralize Omicron. We synthesized codon-optimized spike expression plasmids for Omicron and Delta spike proteins and generated pseudotyped virus (PV) particles by co-transfecting the spike expression plasmids with a lentiviral gag-pol-expressing plasmid and a lentiviral transduction plasmid encoding the luciferase gene,. We obtained longitudinal serum samples from 40 individuals who were vaccinated with either the BNT162b2 or ChAdOx-1 vaccine and performed serum titrations before mixing sera with our reporter PV particles. The participants had a median age of around 70 years and prospective serum samples were taken as follows: one month after dose two, six months after dose two and one month after dose three (Extended Data Table ). We observed at least tenfold loss of neutralization against Omicron after the s..."
},
{
"@type": "HowToStep",
"position": 3,
"name": "Omicron and polyclonal antibodies",
"text": "To confirm the loss of neutralizing activity against Omicron after the second dose, we next used a live virus experimental system to compare Delta and Omicron variants against sera taken four weeks after the second dose of BNT162b2, and obtained similar results to those obtained in the PV assay (Extended Data Fig. ). These live viruses were also used to assess the neutralization of the Omicron variant by sera derived from unvaccinated individuals who were previously infected with the early Wuhan-Hu-1 virus or the Delta variant. As expected, vaccine sera had significantly impaired activity against Omicron compared with Delta (Extended Data Fig. ). We also tested mRNA-1273 vaccine-elicited sera, which showed a similar reduction in neutralization to BNT162b2. However, coronavac sera showed little neutralization against Delta and 0 out of 9 participants had detectable neutralization again..."
},
{
"@type": "HowToStep",
"position": 4,
"name": "Omicron replication and spike cleavage",
"text": "We infected primary human nasal epithelial 3D cultures (hNECs) with an air-liquid interface (Fig. ). Infection with live SARS-CoV-2 Omicron or Delta was conducted at the apical surface with equal amounts of input virus and virus collected from the apical surface at 24 and 48 h, and virus was quantified using quantitative PCR (qPCR) analysis of the E gene and 50% tissue culture infectious dose (TCID 50 ). We observed similar replication kinetics for Omicron and Delta in the hNECs (Fig. ). We next infected Calu-3 lung cells (which are known to express endogenous TMPRSS2 ) with live isolates, and observed significantly greater viral replication for Delta than Omicron (Fig. ), manifesting as early as 24 h after infection. We also infected Caco-2 cells, a colon cancer cell line (known to express high levels of endogenous TMPRSS2 ) and, as for Calu-3, we found 1 log..."
},
{
"@type": "HowToStep",
"position": 5,
"name": "Impaired cell-cell fusion by Omicron S",
"text": "The ability of viral membrane glycoproteins to induce cell-cell fusion and syncytium formation is well established -, providing an additional route for SARS-CoV-2 dissemination that may facilitate evasion of neutralizing antibodies. The role of syncytium formation in viral replication and the pathogenesis of severe COVID-19 has been reported and may be a druggable process to treat COVID-19 pathology,. SARS-CoV-2-mediated cell-cell fusion requires the PBCS and spike cleavage at S1/S2, and the process is known to be accelerated by the presence of TMPRSS2 (ref. )."
},
{
"@type": "HowToStep",
"position": 6,
"name": "Impaired cell-cell fusion by Omicron S",
"text": "Given the requirement of TMPRSS2 for optimal cell-cell fusion, we hypothesized that Omicron may be impaired in mediating this process. We used a split GFP system to monitor cell-cell fusion in real time (Fig. ). As a control to show that spike cleavage is needed for fusion in our assay system, we titrated CMK (a furin inhibitor) into donor cell medium before transfection with spike-expressing plasmids, observing dose-dependent inhibition of fusion (Extended Data Fig. ). As a further control to demonstrate the need for ACE2 engagement by expressed spike, we tested the ability of convalescent serum containing neutralizing antibodies to block cell-cell fusion. Indeed, serum blocked syncytia formation in a dose-dependent manner (Extended Data Fig. ). Fig. 4 SARS-CoV-2 Omicron variant spike shows impaired cell-cell fusion activity and smaller infection foci generate..."
},
{
"@type": "HowToStep",
"position": 7,
"name": "Impaired cell-cell fusion by Omicron S",
"text": "We predicted that poor fusion would impair cell-cell spread of Omicron, and analysed infection focus size using live virus infection of H1299 cells, in which we have shown similar entry efficiency for Omicron and Delta spike PV (Fig. ). Infection foci in spreading virus infections occur because of localized (cell-to-cell) infection, probably facilitated by syncytium formation. The infection was followed by overlay of semi-solid medium (carboxymethylcellulose) to inhibit cell-free infection, thereby favouring cell-cell infection. Infected cells were stained with an anti-S antibodies, and local spread was characterized by the staining of foci visible to the naked eye (Fig. ). Omicron infection resulted in slightly higher numbers of foci (Fig. ), but each infection focus was substantially smaller in size relative to those formed by Delta infection (Fig. ). For a more detailed..."
},
{
"@type": "HowToStep",
"position": 8,
"name": "Flow cytometry analysis of spike protein surface expression",
"text": "ExpiCHO cells were seeded into 50 ml Mini Bioreactor Centrifuge Tube (Corning) at 6 × 10 6 cells per ml in 5 ml ExpiCHO Expression medium (Life technology). Plasmids encoding SARS-CoV-2 Wuhan, Alpha (B.1.1.7), Delta (B.1.617.2) or Omicron (B.1.1.529) spike protein (5 mg) were diluted in OptiPRO SFM (Life Technology) and mixed with Expifectamine CHO Reagent (Life Technology). After incubation for 1 min at room temperature, transfection mixes were added to the cell suspensions. Next, the cells were incubated at 37 °C under 8% CO 2 with an orbital shaking speed of 120 rpm for the next 48 h."
}
],
"tool": [],
"supply": [],
"isBasedOn": {
"@type": "ScholarlyArticle",
"headline": "Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts infectivity and fusogenicity",
"datePublished": "2022",
"author": [
{
"@type": "Person",
"name": "Bo Meng"
},
{
"@type": "Person",
"name": "Adam Abdullahi"
},
{
"@type": "Person",
"name": "Isabella A. T. M. Ferreira"
},
{
"@type": "Person",
"name": "Niluka Goonawardane"
},
{
"@type": "Person",
"name": "Akatsuki Saito"
},
{
"@type": "Person",
"name": "Izumi Kimura"
},
{
"@type": "Person",
"name": "Daichi Yamasoba"
},
{
"@type": "Person",
"name": "Pehuén Pereyra Gerber"
},
{
"@type": "Person",
"name": "Saman Fatihi"
},
{
"@type": "Person",
"name": "Surabhi Rathore"
},
{
"@type": "Person",
"name": "Samantha K. Zepeda"
},
{
"@type": "Person",
"name": "Guido Papa"
},
{
"@type": "Person",
"name": "Steven A. Kemp"
},
{
"@type": "Person",
"name": "Terumasa Ikeda"
},
{
"@type": "Person",
"name": "Mako Toyoda"
},
{
"@type": "Person",
"name": "Toong Seng Tan"
},
{
"@type": "Person",
"name": "Jin Kuramochi"
},
{
"@type": "Person",
"name": "Shigeki Mitsunaga"
},
{
"@type": "Person",
"name": "Takamasa Ueno"
},
{
"@type": "Person",
"name": "Kotaro Shirakawa"
},
{
"@type": "Person",
"name": "Akifumi Takaori-Kondo"
},
{
"@type": "Person",
"name": "Teresa Brevini"
},
{
"@type": "Person",
"name": "Donna L. Mallery"
},
{
"@type": "Person",
"name": "Oscar J. Charles"
},
{
"@type": "Person",
"name": "Stephen Baker"
},
{
"@type": "Person",
"name": "Gordon Dougan"
},
{
"@type": "Person",
"name": "Christoph Hess"
},
{
"@type": "Person",
"name": "Nathalie Kingston"
},
{
"@type": "Person",
"name": "Paul J. Lehner"
},
{
"@type": "Person",
"name": "Paul A. Lyons"
},
{
"@type": "Person",
"name": "Nicholas J. Matheson"
},
{
"@type": "Person",
"name": "Willem H. Ouwehand"
},
{
"@type": "Person",
"name": "Caroline Saunders"
},
{
"@type": "Person",
"name": "Charlotte Summers"
},
{
"@type": "Person",
"name": "James E. D. Thaventhiran"
},
{
"@type": "Person",
"name": "Mark Toshner"
},
{
"@type": "Person",
"name": "Michael P. Weekes"
},
{
"@type": "Person",
"name": "Patrick Maxwell"
},
{
"@type": "Person",
"name": "Ashley Shaw"
},
{
"@type": "Person",
"name": "Ashlea Bucke"
},
{
"@type": "Person",
"name": "Jo Calder"
},
{
"@type": "Person",
"name": "Laura Canna"
},
{
"@type": "Person",
"name": "Jason Domingo"
},
{
"@type": "Person",
"name": "Anne Elmer"
},
{
"@type": "Person",
"name": "Stewart Fuller"
},
{
"@type": "Person",
"name": "Julie Harris"
},
{
"@type": "Person",
"name": "Sarah Hewitt"
},
{
"@type": "Person",
"name": "Jane Kennet"
},
{
"@type": "Person",
"name": "Sherly Jose"
},
{
"@type": "Person",
"name": "Jenny Kourampa"
},
{
"@type": "Person",
"name": "Anne Meadows"
},
{
"@type": "Person",
"name": "Criona O'Brien"
},
{
"@type": "Person",
"name": "Jane Price"
},
{
"@type": "Person",
"name": "Cherry Publico"
},
{
"@type": "Person",
"name": "Rebecca Rastall"
},
{
"@type": "Person",
"name": "Carla Ribeiro"
},
{
"@type": "Person",
"name": "Jane Rowlands"
},
{
"@type": "Person",
"name": "Valentina Ruffolo"
},
{
"@type": "Person",
"name": "Hugo Tordesillas"
},
{
"@type": "Person",
"name": "Ben Bullman"
},
{
"@type": "Person",
"name": "Benjamin J. Dunmore"
},
{
"@type": "Person",
"name": "Stefan Gräf"
},
{
"@type": "Person",
"name": "Josh Hodgson"
},
{
"@type": "Person",
"name": "Christopher Huang"
},
{
"@type": "Person",
"name": "Kelvin Hunter"
},
{
"@type": "Person",
"name": "Emma Jones"
},
{
"@type": "Person",
"name": "Ekaterina Legchenko"
},
{
"@type": "Person",
"name": "Cecilia Matara"
},
{
"@type": "Person",
"name": "Jennifer Martin"
},
{
"@type": "Person",
"name": "Federica Mescia"
},
{
"@type": "Person",
"name": "Ciara O'Donnell"
},
{
"@type": "Person",
"name": "Linda Pointon"
},
{
"@type": "Person",
"name": "Joy Shih"
},
{
"@type": "Person",
"name": "Rachel Sutcliffe"
},
{
"@type": "Person",
"name": "Tobias Tilly"
},
{
"@type": "Person",
"name": "Carmen Treacy"
},
{
"@type": "Person",
"name": "Zhen Tong"
},
{
"@type": "Person",
"name": "Jennifer Wood"
},
{
"@type": "Person",
"name": "Marta Wylot"
},
{
"@type": "Person",
"name": "Ariana Betancourt"
},
{
"@type": "Person",
"name": "Georgie Bower"
},
{
"@type": "Person",
"name": "Chiara Cossetti"
},
{
"@type": "Person",
"name": "Aloka De Sa"
},
{
"@type": "Person",
"name": "Madeline Epping"
},
{
"@type": "Person",
"name": "Stuart Fawke"
},
{
"@type": "Person",
"name": "Nick Gleadall"
},
{
"@type": "Person",
"name": "Richard Grenfell"
},
{
"@type": "Person",
"name": "Andrew Hinch"
},
{
"@type": "Person",
"name": "Sarah Jackson"
},
{
"@type": "Person",
"name": "Isobel Jarvis"
},
{
"@type": "Person",
"name": "Ben Krishna"
},
{
"@type": "Person",
"name": "Francesca Nice"
},
{
"@type": "Person",
"name": "Ommar Omarjee"
},
{
"@type": "Person",
"name": "Marianne Perera"
},
{
"@type": "Person",
"name": "Martin Potts"
},
{
"@type": "Person",
"name": "Nathan Richoz"
},
{
"@type": "Person",
"name": "Veronika Romashova"
},
{
"@type": "Person",
"name": "Luca Stefanucci"
},
{
"@type": "Person",
"name": "Mateusz Strezlecki"
},
{
"@type": "Person",
"name": "Lori Turner"
}
],
"identifier": "10.1038/s41586-022-04474-x"
}
},
{
"@context": "https://schema.org",
"@type": "BreadcrumbList",
"itemListElement": [
{
"@type": "ListItem",
"position": 1,
"name": "Experiments",
"item": "https://replicatescience.com/experiments"
},
{
"@type": "ListItem",
"position": 2,
"name": "Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts infectivity and fusogenicity methods",
"item": "https://replicatescience.com/experiments/altered-tmprss2-usage-by-sars-cov-2-omicron-impacts-infectivity-and-fusogenicity-methods-bo-meng-pmc8942856/altered-tmprss2-usage-by-sars-cov-2-omicron-impacts-infectivity-and-fusogenicity-mlph26w8"
}
]
}
]