Cat.No. : CSC-RT0101
Host Cell: HEK293 Target Gene: SLC1A5
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
| Cat. No. | CSC-RT0101 |
| Description | This cell line is a stable cell line with a homozygous knockout of human SLC1A5 using CRISPR/Cas9. |
| Target Gene | SLC1A5 |
| Gene ID | 6510 |
| Genotype | SLC1A5 (-/-) |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Cell Type | Epithelial |
| Size | 1x10^6 cells/vial, 1mL |
| SequencingResult | Homozygous: 34 bp deletion in exon |
| 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 25cm2 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 |
Alanine-serine-cysteine transporter 2 (ASCT2, SLC1A5) is the major transporter of glutamine in cancer cells. In this study, researchers developed a novel monoclonal antibody (mAb) that recognizes the extracellular domain of human ASCT2 and investigated whether ASCT2 could be a therapeutic target for KRAS mutant cancers. Rats were immunized with RH7777 rat hepatoma cells expressing human ASCT2 fused to green fluorescent protein (GFP). Splenocytes from immunized rats were fused with P3X63Ag8.653 mouse myeloma cells, and hybridoma cells secreting Ab3-8 mAb were established for screening and cloning. The mAb reacted with RH7777 transfectants expressing ASCT2-GFP protein in a GFP intensity-dependent manner. Ab3-8 reacted with various human cancer cells but not with noncancerous mammary epithelial cells or ASCT2 knockout HEK293 and SW1116 cells. In SW1116 and HCT116 human colon cancer cells with KRAS mutations, treatment with Ab3-8 reduced intracellular glutamine transport, phosphorylation of AKT and ERK, and inhibited tumor growth of these cells in athymic mice. Ab3-8 inhibition of in vivo tumor growth was not observed in HT29 colon and HeLa uterine cancer cells with wild-type KRAS. These results suggest that ASCT2 is an excellent therapeutic target for KRAS mutant cancers.
Ab3-8 did not react with ASCT2 (SLC1A5) Knockout Cell Line-HEK293 (ASCT2-KO-HEK293) and SW1116 cells (Figure 1E). In addition, Ab3-8 reacted with various human cancer cells but was weakly reactive with non-cancerous cells (Figure 1F). Among these cell lines, SW1116, HCT116, DLD-1, and HCT15 cells contained KRAS mutations, while the other cell lines had wild-type KRAS.
Figure 1. E, ASCT2-KO HEK293 and SW1116 cells were reacted with Ab3-8 and subsequently with PE-conjugated anti-rat IgG. The reactivity of Ab3-8 with these cells was analyzed by flow cytometry. F, FCM analysis of cell surface expression of ASCT2 protein in various human cell lines. (Hara, Yuta, et al. 2020)
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.
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We are studying the effects of SLC1A5 knockout on cancer cell growth and proliferation. This cell line is very helpful for our research.
Great buy! The SLC1A5 knockout cells lack functional SLC1A5 protein. Good experimental results were obtained. I recommend it.
I am extremely impressed with the effectiveness of SLC1A5 Knockout Cell Line-HEK293. Its high knockout efficiency and reliable results have allowed me to confidently investigate the role of specific genes in biological processes.
SLC1A5 Knockout Cell Line-HEK293 has significantly enhanced the precision and accuracy of my experiments. Its robust performance and reproducible knockout outcomes have provided me with reliable data for my research.
Using SLC1A5 Knockout Cell Line-HEK293 has been a distinguished assistants for my studies. Its user-friendly interface, combined with its exceptional knockout efficiency, has made gene editing experiments more efficient and successful.
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