Cat.No. : CSC-RT1517
Host Cell: HeLa Target Gene: PTGS2
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
| Cat. No. | CSC-RT1517 |
| Description | A stable cell line with a homozygous knockout of human PTGS2 using CRISPR/Cas9. |
| Target Gene | PTGS2 |
| Host Cell | HeLa |
| Host Cell Species | Homo sapiens (Human) |
| Shipping | 10^6 cells/tube |
| Storage | Liquid nitrogen |
| 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 1:4-1:6. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| Gene Name | PTGS2 |
| Gene ID | 5743 |
| 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 |
Cytotoxic therapies, in addition to directly inducing cancer cell death, can stimulate immune-dependent tumor growth control or accelerate tumor progression. Here, researchers show that cytotoxic therapies acutely upregulate cyclooxygenase (COX)-2 (also known as Prostaglandin-endoperoxide synthase 2 (PTGS2)) expression and prostaglandin E2 (PGE2) production in cancer cells with pre-existing COX-2 activity. Screening of a compound library of 1280 approved drugs revealed that chemotherapeutic agents from all classes enhance COX-2 transcription while inhibiting cancer cell proliferation. Genetic manipulation of COX-2 expression or its gene promoter region revealed how enhanced COX-2/PGE2 activity after treatment profoundly alters the inflammatory properties of chemotherapy-treated cancer cells in vivo. Pharmacological COX-2 inhibition enhances the efficacy of combined chemotherapy and PD-1 blockade. These findings suggest that COX-2/PGE2 upregulation by dying cancer cells is a major barrier to cytotoxic therapy-driven tumor immunity and reveal a strategy to improve outcomes of combined immunotherapy and chemotherapy treatments.
Here, to assess the contribution of the COX-2/PGE2 axis to the inflammatory phenotype of CTX-treated cells, the researchers compared the effects of injection of cisplatin or 5-FU-treated COX-2 WT and COX-2 (PTGS2) knockout 4T1 cells. Notably, CTX-treated COX-2 knockout cells attracted far fewer neutrophils or monocytes than their COX-2-expressing parental cells and were similar to untreated 4T1 cells (Figure 1), indicating a major contribution of cancer cell-intrinsic COX-2 activity. COX-2 REST cells pre-treated with cisplatin or 5-FU were less effective than COX-2 WT cells in recruiting neutrophils and monocytes and largely phenotypically mimicked CTX-treated COX-2 knockout cells (Figure 1). Together, these results suggest that transcriptional upregulation of COX-2 by cancer cells after CTX, rather than its basal expression, largely explains their ability to stimulate recruitment of inflammatory myeloid cells.
Figure 1. Total number of neutrophils and monocytes recruited by i.p. injection of untreated or CTX-treated 4T1 COX-2 WT, COX-2 knockout (KO), or COX-2 REST cells. (Bell, Charlotte 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.
PTGS2 knockout cells are cells that have been genetically engineered to lack the PTGS2 gene and thus do not express the PTGS2 protein. This PTGS2 knockout cell line is very helpful for our research.
The use of PTGS2 knockout cells has aided in the development of highly selective PTGS2 inhibitors. I recommend Creative Biogene'cell line.
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