Human ACE2 Knockout Cell Line-HEK293T
Cat.No. : CSC-RT2688
Host Cell: HEK293T Target Gene: ACE2
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2688 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of human ACE2 using CRISPR/Cas9. |
| Target Gene | ACE2 |
| Host Cell | HEK293T |
| Size Form | 1 vial (10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid nirtogen |
| Species | Human |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:3-1:6. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
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Background
Case Study
Applications
Angiotensin-converting enzyme 2 (ACE2) is a key protein in the renin-angiotensin system (RAS) that plays a critical role in cardiovascular health and fluid balance. ACE2 is primarily found on the surface of various cell types, including alveolar epithelial cells, small intestinal enterocytes, endothelial cells, and arterial smooth muscle cells. It has several critical physiological functions, including converting angiotensin II, a potent vasoconstrictor, into angiotensin-(1-7), which promotes vasodilation and has anti-inflammatory properties.
ACE2 has received extensive attention during the COVID-19 pandemic because it is a receptor for the SARS-CoV-2 virus. The viral spike protein binds to ACE2 to facilitate entry into host cells. The widespread expression of ACE2 in different tissues explains the multi-organ effects frequently observed in COVID-19 patients. This interaction has prompted extensive research into inhibitors or modulators of ACE2 as potential therapeutic avenues for COVID-19. In addition to COVID-19, ACE2 remains a focus of research in hypertension, heart failure, and kidney disease. Understanding the dynamics of ACE2 within the RAS and its broader impact may lead to new treatments for a variety of cardiovascular and inflammatory diseases. ACE2 therefore remains a protein of great biomedical importance, impacting multiple areas of health and disease.
Studies have shown that SARS-CoV-2 infects human monocytes, monocyte-derived macrophages, and dendritic cells in vitro, which may play an important role in the pathogenesis of COVID-19. However, whether SARS-CoV-2 can infect lymphocytes that do not express ACE2 and cause lymphocytopenia remains unknown. Here, the study showed that activated T lymphocytes can be infected by SARS-CoV-2 in an ACE2-independent manner. This infection led to significant apoptosis of T cells in vitro or in COVID-19 patients. The results of this study help understand lymphocytopenia caused by SARS-CoV-2 infection.
ACE2 is widely believed to be the entry receptor for SARS-CoV-2. However, the major cell population in peripheral blood cells (PBCs) expressed extremely low levels of ACE2, raising the question of whether ACE2 also mediates SARS-CoV-2 viral entry into T cells. The researchers first tested whether ACE2 knockdown could inhibit SARS-CoV-2 infection of T cells. The data showed that ACE2 was successfully knocked down by ACE2-shRNA in Caco2 cells. Jurkat T cells did not express detectable ACE2 under either mock or knockdown conditions (Figure 1a). Correspondingly, ACE2 knockdown resulted in a significant reduction in SARS-CoV-2 infection in Caco2 cells, but had no such effect in Jurkat T cells (Figure 1b). To further confirm this finding, ACE2 was knocked out in Caco2 and Jurkat cells (Figure 1c). Similar to ACE2 knockdown cells, the viral load decreased in ACE2 knockout cells-Caco2, but not in ACE2 knockout cells-Jurkat (Figure 1d). These results suggest that SARS-CoV-2 infects T cells in an ACE2-independent manner.
Figure 1. SARS-CoV-2 infection of T cell is spike-ACE2/TMPRSS2-independent. (Shen X R, et al., 2022)
The Human ACE2 Knockout Cell Line-HEK293T offers numerous applications in biomedical research, particularly in virology, drug discovery, and molecular biology. Below are some primary applications:
Viral Entry Mechanism Investigation: HEK293T ACE2 knockout cells serve as a critical tool for studying the mechanisms of SARS-CoV-2 entry, as ACE2 is the primary receptor for the virus. By comparing knockout cells to wild-type cells, researchers can gain insights into alternative pathways or co-receptors involved in viral infection.
Antiviral Screening: They are used to screen antiviral compounds and neutralizing antibodies targeting different steps in the viral lifecycle, specifically those that may inhibit the interaction between the virus and the ACE2 receptor.
Functional Analysis of ACE2: Using these knockout cell lines, researchers can study the physiological role of ACE2 under normal and pathological conditions, including its role in cardiovascular diseases, hypertension, and other ACE2-related functions.
Vaccine Development: These cell lines can be used for validating the efficacy of vaccines that target the ACE2-SARS-CoV-2 interaction, helping to ensure that vaccine candidates effectively block this critical interaction in a human cell context.
High-Throughput Screening: Utilize the knockout cell line in high-throughput screens to discover novel therapeutic targets and compounds.
For research use only. Not intended for any clinical use.
