Cat.No. : CSC-RT2701
Host Cell: HCT116 Target Gene: MTAP
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
| Cat. No. | CSC-RT2701 |
| Description | This cell is a stable cell line with a homozygous knockout of human MTAP using CRISPR/Cas9. |
| Target Gene | MTAP |
| Host Cell | HCT116 |
| 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. |
| 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 |
The methylthioadenosine phosphorylase (MTAP) gene is located adjacent to the cyclin-dependent kinase inhibitor 2A (CDKN2A) tumor suppressor gene and is co-deleted with CDKN2A in approximately 15% of cancers. This co-deletion leads to malignancy, poor prognosis, and lack of effective molecular targeted therapies. The metabolic enzyme methionine adenosyltransferase 2α (MAT2A) has been identified as a synthetic lethal target in MTAP-deficient cancers. Here, researchers report the characterization of potent MAT2A inhibitors that significantly reduce S-adenosylmethionine (SAM) levels and exhibit antiproliferative activity in MTAP-deficient cancer cells and tumors. Using RNA sequencing and proteomics, they demonstrate that MAT2A inhibition is mechanistically linked to reduced protein arginine methyltransferase 5 (PRMT5) activity and splicing perturbations. We further show that MAT2A inhibition leads to DNA damage and mitotic defects in HCT116 MTAP knockout cells, providing a rationale for combining the MAT2A clinical candidate AG-270 with antimitotic taxanes.
AGI-24512, a MAT2A inhibitor, showed 3-6 orders of magnitude greater cellular potency than previously reported MAT2A inhibitors PF-9366 and cycloleucine. Reduced incorporation of thethymidine analog 5-ethynyl-2′-deoxyuridine (EdU) was measured by immunofluorescence analysis in MTAP knockout (MTAP−/−) cells but not in WT cells (Figures 1A and 1B). However, no significant cell cycle checkpoint activation was detected after inhibition of MAT2A in asynchronously proliferating cells. As AGI-24512-induced cell cycle alterations may be more pronounced in synchronized cells, a double thymidine blockade was performed to synchronize cells pretreated with AGI-24512 for 72 hours in early S phase (Figure 1C). After release from replication block, progression from G1 into S and G2/M phases was significantly attenuated in MTAP−/− cells treated with AGI-24512, but not in WT cells (Figures 4C and 4D). These cell cycle changes were accompanied by DNA damage at the chromosomal level, as indicated by the formation of micronuclei and the appearance of binucleated and multinucleated cells after treatment with AGI-24512 (Figures 1E-1H). Both the formation of micronuclei and the appearance of multinucleated cells were restricted to MTAP−/− cells and absent in WT cells.
Figure 1. Pharmacologic Inhibition of MAT2A Leads to MTAP−/− Genotype-Selective Cell-Cycle Delays and Mitotic Defects. (Kalev P, et al., 2021)
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.
The cells arrived quickly and in perfect condition, allowing us to dive straight into our experiments. We no longer have to spend precious weeks generating knockout cell lines ourselves.
As a researcher studying cancer metabolism, the Human MTAP Knockout Cell Line-HCT116 has significantly enhanced our ability to explore therapeutic targets.
Write a review of your use of Biogene products and services in your research. Your review can help your fellow researchers make informed purchasing decisions.
Our promise to you:
Guaranteed product quality, expert customer support.
24x7 CUSTOMER SERVICE
CONTACT US TO ORDER
Copyright © Creative Biogene. All rights reserved.
