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Human ACE2 293T Cell Line

Entry of SARS-CoV-2 into host cells is mediated by physical interaction between viral spike proteins and ACE2

Entry of SARS-CoV-2 into host cells is mediated by interaction between spike proteins and ACE2. To aid the study of virus-receptor interactions and the development of therapies for COVID-19, we have developed a stable HEK-derived cell line that expresses ACE2 under the CMV promoter. The cell line has been confirmed to express ACE2 at consistently high levels across multiple passages and is efficiently transduced by SARS-CoV-2 pseudovirus generated using Lenti-X SARS-CoV-2 Packaging Single Shots.

Entry of SARS-CoV-2 into host cells is mediated by interaction between spike proteins and ACE2. To aid the study of virus-receptor interactions and the development of therapies for COVID-19, we have developed a stable HEK-derived cell line that expresses ACE2 under the CMV promoter. The cell line has been confirmed to express ACE2 at consistently high levels across multiple passages and is efficiently transduced by SARS-CoV-2 pseudovirus generated using Lenti-X SARS-CoV-2 Packaging Single Shots.

ACE2 is a zinc-containing metalloenzyme located on the surface of endothelial and other cell types. This transmembrane protein converts angiotensin I to angiotensin 1–9, a 9-amino acid peptide with anti-hypertrophic effects in cardiomyocytes, and angiotensin II to angiotensin 1–7, which then acts as a beneficial vasodilator and anti-proliferation agent.

The Human ACE2 293T Cell Line was engineered by transduction of an HEK cell line with lentivirus encoding human ACE2. The cell line was subcloned for high expression of ACE2, and confirmed via RT-qPCR.

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Cat. # Product Size Price License Quantity Details
631289 Human ACE2 293T Cell Line 1 mL USD $1950.00

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ID Number  
406 This product is the subject of a technology license agreement for internal research use only. Use of this product other than for research use may require additional licenses. Information on license restrictions or for uses other than research may be obtained by contacting licensing@takarabio.com.

The Human ACE2 293T Cell Line is a transduced human embryonic kidney-derived cell line that constitutively expresses the human angiotensin I converting enzyme 2 (ACE2) under the CMV promoter. The cell line has been confirmed by RT-qPCR to express ACE2 at consistently high levels across multiple passages, and shown to be efficiently transduced by SARS-CoV-2 pseudovirus.

Cat. # 631289 consists of a tube containing 2 x 106 cells.

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Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Analysis of ACE2 expression by flow cytometry

Analysis of ACE2 expression by flow cytometry

Analysis of ACE2 expression by flow cytometry. ACE2 expression at the cell surface of the Human ACE2 293T Cell Line was detected by flow cytometry using an Alexa Fluor 488-conjugated monoclonal anti-ACE2 antibody (R&D Systems, Cat. # FAB9332G). Non-labelled cells were used as a negative control.

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Confirmation of stable ACE2 expression across multiple cell passages

Confirmation of stable ACE2 expression across multiple cell passages

Confirmation of stable ACE2 expression across multiple cell passages. Relative mRNA expression levels of ACE2 across successive passages of the Human ACE2 293T Cell Line were determined by RT-qPCR using the One Step PrimeScript RT-PCR Kit (Perfect Real Time) (Cat. # RR064A). Ct values for ACE2 expression were normalized to Ct values for the housekeeping gene ACTB in triplicates. ACE2 expression in the stable cell line was determined relative to a 293T cell line by calculating 2^(–ΔΔCt).

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Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (truncated form) and encoding the fluorescent protein ZsGreen1. 100 μl of supernatant from each prep was used to transduce the Human ACE2 293T Cell Line. Panel A. Microscopy images of transduced cells at 72 hours post-infection. Panel B. Transduction efficiencies for each sample were measured by flow cytometry 6 days post-transduction.

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Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

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Neutralization data with convalescent serum

Neutralization data with convalescent serum

Use of the Human ACE2 293T Cell Line in neutralization assays employing pooled non-human primate convalescent serum to SARS-CoV-2 as a blocking agent. WT (truncated) SARS-CoV-2 pseudovirus was incubated with serial dilutions of the convalescent serum (BEI Resources, Cat. # NR-52401) and applied to the Human ACE2 293T Cell Line. Luciferase levels were measured 3 days post-infection as a readout for virus infectivity. Data are graphed as percent neutralization relative to virus-only control infection. Values are mean ±SD and experiments were performed in triplicate. Negative control was performed using serum from healthy macaques.

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Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. Panels A and B. Serial dilutions of soluble ACE2 protein fused to the Fc domain from IgG (ACE2-Fc) were applied, along with SARS-CoV-2 pseudovirus, to the Human ACE2 293T Cell Line. Luciferase levels were measured 3 days post-infection as a readout for virus infectivity. Data are graphed as percent neutralization relative to virus-only control infection. Values are mean ±SD and experiments were performed in triplicate.

SARS-CoV-2 pseudovirus
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Overview

  • HEK cell line engineered for constitutive expression of ACE2 under the CMV promoter
  • Consistently high expression of ACE2, confirmed across multiple passages
  • Efficiently transduced by SARS-CoV-2 pseudovirus (e.g., produced using Lenti-X SARS-CoV-2 Packaging Single Shots)

More Information

Applications

  • SARS-CoV-2 neutralization studies
  • Analysis of virus-receptor interactions and viral entry

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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