Transfected Stable Cell Lines
Reliable | High-Performance | Wide Rage
Precision reporter, kinase, immune receptor, biosimilar, Cas9, and knockout stable cell lines for diverse applications.
Cat. No. : CSC-RT0518
Target Gene : KRAS Host Cell : HeLa
Size : >1x106 cells/vial Validation : Sequencing
| Cat. No. | CSC-RT0518 |
| Description | This cell line is a stable cell line with a homozygous knockout of human KRAS using CRISPR/Cas9. |
| Cell Type | Epithelial |
| Target Gene | KRAS |
| Host Cell | HeLa |
| Host Cell Species | Homo sapiens (Human) |
| Genotype | KRAS (-/-) |
| Size | >1x10^6 cells/vial, 1 mL |
| Validation | Sequencing |
| Sequencing Result | Homozygous: 1 bp insertion 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 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 |
| 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 |
| Gene Name | KRAS v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog [ Homo sapiens ] |
| Gene Symbol | KRAS |
| Synonyms | NS; NS3; KRAS1; KRAS2; RASK2; KI-RAS; C-K-RAS; K-RAS2A; K-RAS2B; K-RAS4A; K-RAS4B |
| Gene Description | v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog |
| Gene ID | 3845 |
| Uni Prot ID | P01116 |
| m RNA Refseq | NM_033360.2 |
| Protein Refseq | NP_203524.1 |
| Chromosome Location | 12p12.1 |
| Function | GDP binding; GMP binding; GTP binding; GTPase activity; LRR domain binding; protein binding; |
| Pathway | ARMS-mediated activation, organism-specific biosystem; Activation of RAS in B Cells, organism-specific biosystem; Acute myeloid leukemia, organism-specific biosystem; Acute myeloid leukemia, conserved biosystem; Adaptive Immune System, organism-specific biosystem; Alcoholism, organism-specific biosystem; Alcoholism, conserved biosystem; |
| MIM | 190070 |
KRAS (Kirsten rat sarcoma viral oncogene homolog) is a gene that encodes a protein involved in cellular signaling. As part of the RAS oncogene family, the KRAS gene is related to the HRAS and NRAS genes and is an integral part of the MAPK/ERK signaling pathway, which mediates various cellular functions, including proliferation, survival, and growth.
Mutations in the KRAS gene are known to be associated with various types of cancer, making it one of the most studied genes in oncology. KRAS mutations are particularly common in pancreatic, colorectal, and lung cancers. In pancreatic ductal adenocarcinoma (PDAC), nearly 90% of cases exhibit KRAS mutations, making it a key target for therapeutic intervention. Similarly, approximately 40% of colorectal cancers and 15-25% of lung adenocarcinomas carry KRAS mutations. Despite the importance of KRAS, direct targeting of KRAS has been challenging due to the protein’s high affinity for GTP/GDP and lack of binding sites suitable for drug design.
Recent advances show promise, particularly the development of KRAS G12C inhibitors, which specifically target the mutant form of the protein found in certain cancers. Drugs such as sotorasib and adagrasib have shown efficacy in treating patients with KRAS G12C mutant non-small cell lung cancer, marking a major breakthrough in precision oncology.
Human KRAS knockout HeLa cell lines are valuable tools in biomedical research. Here are a few key applications of human KRAS knockout HeLa cell lines:
Cancer Research: The KRAS gene is frequently mutated in various cancers, making the KRAS knockout HeLa cell line valuable for studying oncogenic mechanisms. Researchers can study how loss of KRAS affects cell signaling, tumor growth, and metastasis, providing insights into potential therapeutic targets.
Drug Screening: These cell lines are an important tool for high-throughput screening of anticancer drugs. By evaluating the efficacy and toxicity of new compounds in KRAS knockout HeLa cells, scientists can identify drug candidates that selectively target KRAS mutant pathways.
Gene Function Studies: KRAS knockout HeLa cells enable researchers to study the role of KRAS in cellular processes such as cell proliferation, differentiation, and apoptosis.
Signal Transduction Studies: KRAS is a key player in the RAS/MAPK signaling pathway. Using knockout cell lines, researchers can dissect the complexity of the pathway and determine how KRAS interacts with other proteins and contributes to signaling, which is critical for developing pathway-specific inhibitors.
Study of Resistance Mechanisms: Cancer cells often become resistant to therapies. By using KRAS knockout HeLa cells, researchers can explore how the loss of KRAS affects resistance mechanisms.
A: DMEM supplemented with 10% fetal bovine serum. <br> 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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