Cat.No. : CSC-RT2688
Host Cell: HEK293T Target Gene: ACE2
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
| Cat. No. | CSC-RT2688 |
| Description | This cell is a stable cell line with a homozygous knockout of human ACE2 using CRISPR/Cas9. |
| Target Gene | ACE2 |
| Host Cell | HEK293T |
| Host Cell Species | Homo sapiens (Human) |
| Size Form | 1 vial (10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid nirtogen |
| 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 |
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)
A: DMEM supplemented with 10% fetal bovine serum.
It is not required to add the selection antibiotics when culturing the KO cells.
A: The knockout cell product is validated by PCR amplification and Sanger Sequencing to confirm the mutation at the genomic level. Please find the detailed mutation info in the datasheet.
A: Single clonal cell.
A: No. This knockout cell product is generated using the CRISPR/Cas9 system to induce small insertions or deletions (indels) resulting in frameshift mutations. Although these frameshift mutations typically disrupt the coding gene, there is a possibility that the non-functional transcript may still be transcribed. Consequently, this could potentially yield misleading results when analyzed by RT-qPCR.
A: The cell line should be stored in liquid nitrogen for long-term preservation.
A: For most cases, we often keep at least 2 clones with different frameshift mutations. Please feel free to contact us to check if there are additional available clones.
If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.
Human ACE2 Knockout Cell Line - HEK293T is a product that allows for relatively efficient knockout of the human ACE2 gene, and the results can be observed with a variety of assays.
Human ACE2 Knockout Cell Line-HEK293T is critical for our studies on cell signaling pathways. The knockout is effective, and the cells are highly viable and easy to culture.
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