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HEK293 NF-κB-reporter cells expressing the SARS-CoV-2 receptors

InvivoGen offers human embryonic kidney 293 (HEK-293)-derived cell lines, specifically designed for COVID-19 studies:

— HEK-Blue™ hACE2-TMPRSS2 cells

— HEK-Blue™ hACE2 cells

These cells have been engineered to stably overexpress the host SARS-CoV-2 receptors, human (h)ACE2, and TMPRSS2 [1]. They were generated from HEK‑Blue™ Null1‑v cells, which express an NF-κB inducible SEAP (secreted embryonic alkaline phosphatase) reporter gene.

More details

Studying infection and cell fusion with
HEK-Blue™ hACE2-TMPRSS2 cells

Available upon request:

• HEK-Lucia™ hACE2 Cells
• HEK-Lucia™ hACE2-TMPRSS2 Cells
• HEK-hACE2 Cells
• HEK-hACE2-TMPRSS2 Cells

Sensitive to SARS-CoV-2

SARS-CoV-2, the causative agent of coronavirus disease-19 (COVID-19), gains entry into host cells through the interaction of the viral Spike protein with the host ACE2 and TMPRSS2 receptors [1]. HEK293-derived cells are poorly permissive to infection by SARS-CoV-2 or Spike pseudotyped lentiviral particles. Therefore, to increase their permissivity HEK-Blue™ Null1-v cells have been stably transfected with ACE2 only, or ACE2 and TMPRSS2. In contrast to HEK-Blue™ Null1-v cells, HEK-Blue™ hACE2(‑TMPRSS2) cells are sensitive to Spike pseudotyped lentiviral particles. Notably, the addition of TMPRSS2 increases the cell line’s infectivity.

For studying Spike-ACE2-dependent cell fusion

These HEK-293-derived SARS-COV-2 permissive reporter cells can be used as ‘acceptor’ cells in InvivoGen’s SARS-CoV-2 Spike-ACE2 dependent cell fusion assay. This relies on the transfer of the TLR/IL-1 adaptor molecule, MyD88, from a 'donor cell line' (293-hMyD88 transfected with a Spike expression plasmid) to an 'acceptor cell line' expressing an NF-κB-SEAP-inducible reporter gene. Cell fusion is readily assessable in the co-culture supernatant using the SEAP detection reagent, QUANTI‑Blue™ Solution. Unlike infection by pseudotyped particles, the presence of TMPRSS2 does not have an effect on cell fusion in this assay.

Key features

Verified overexpression of human ACE2 and TMPRSS2 genes
Permissive and sensitive to SARS-COV-2 Spike pseudotyped lentiviral particles
Functionally tested in InvivoGen’s cell fusion assay with SARS-CoV-2 Spike-expressing hMyD88 cells.
Readily assessable NF-κB-dependent SEAP reporter activity

Applications

Screening inhibitors of Spike-binding and/or its host receptors (i.e. ACE2 and TMPRSS2).
Studying the efficacy of the vaccines and/or current therapeutics (e.g. mAbs) against the emerging variants by infection studies with pseudotyped particles or cell fusion assays.

InvivoGen has also generated:
➔ HEK-Lucia™ hACE2 and HEK-Lucia™ hACE2-TMPRSS2 cells featuring an NF-KB-inducible Lucia luciferase reporter for a larger detection range than the colorimetric SEAP assays.
➔ HEK-hACE2 and HEK-hACE2-TMPRSS2 cells with no reporter activity for infection, pseudoparticle transduction, or qualitative fusion assays.
These cells have been used to compare the efficiency of SARS-CoV2 Spike variant-mediated cellular infection and fusion. See data

Learn more about SARS-CoV-2 infection and potential therapeutics

References

1. Hoffmann M. et al., 2020. SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell. 181:1-16.
2. Zhou P. et al., 2020. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature. 579(7798):270-273.
3. Walls A.C. et al., 2020. Structure, function, and antigenicity of the SARS-CoV-2 spike glycoprotein. Cell. 181(2):281-292.e6.

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