DNMT1 Knockout Cell Line-HEK293T
Cat.No. : CSC-RT0470
Host Cell: HEK293T Target Gene: DNMT1
Size: 1x10^6 cells/vial, 1 mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT0470 |
| Cell Line Information | This cell line is a stable cell line with a homozygous knockout of human DNMT1 gene using CRISPR/Cas9. |
| Target Gene | DNMT1 |
| Gene ID | 1786 |
| Genotype | DNMT1 (-/-) |
| Host Cell | HEK293T |
| Size | 1x10^6 cells/vial, 1 mL |
| 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 1:3-1:6. |
| 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 |
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Background
Applications
DNA (cytosine-5)-methyltransferase 1, commonly referred to as DNMT1, is a key enzyme in mammalian epigenetics. This enzyme plays a critical role in maintaining DNA methylation patterns after DNA replication. The enzyme is ubiquitously expressed but is particularly abundant in proliferating cells, such as those found in embryonic tissues and tumors. Its activity is tightly regulated through interactions with various proteins and post-translational modifications, ensuring that its functions are precisely executed in different cellular contexts. For example, DNMT1 interacts with proliferating cell nuclear antigen (PCNA) during the S phase of the cell cycle, localizing it to replication forks and linking DNA synthesis to the maintenance of methylation.
Mutations and dysregulation of DNMT1 have been implicated in a range of human diseases, including various forms of cancer and neurodevelopmental disorders. Hypomethylation of DNA can lead to genomic instability and activation of oncogenes, whereas hypermethylation can lead to silencing of tumor suppressor genes. Therefore, DNMT1 and its pathway are important targets for therapeutic intervention strategies to treat cancer and other epigenetic diseases. DNA methylation inhibitors such as 5-azacytidine are already in clinical use, and ongoing research will continue to reveal the nuanced roles and mechanisms of DNMT1.
Epigenetic Research: DNMT1 is a key enzyme responsible for maintaining DNA methylation patterns during DNA replication. Knockout of DNMT1 in 293T cells provides a valuable model for studying the dynamics of DNA methylation and its role in the regulation of gene expression. This helps to elucidate the mechanisms underlying epigenetic modifications and their impact on cellular processes such as differentiation, development, and disease progression.
Cancer Research: Aberrant DNA methylation is a hallmark of many cancers. By using DNMT1 knockout 293T cells, researchers can study how DNMT1-mediated loss of methylation affects cellular pathways implicated in tumorigenesis. This model can be used to identify potential biomarkers for cancer diagnosis and prognosis, as well as to screen and evaluate the efficacy of novel anticancer therapies targeting DNA methylation machinery.
Stem Cell Research: DNMT1 plays a key role in maintaining stem cell pluripotency and differentiation. The 293T DNMT1 knockout cell line can be used to study the role of DNA methylation in stem cell biology, including maintenance of stem cell identity and regulation of lineage commitment.
Pharmacological Testing: The DNMT1 knockout 293T cell line is a valuable tool for testing compounds that modulate DNA methylation. It can be used to evaluate the effects of DNMT inhibitors and other epigenetic drugs, providing a platform for drug discovery and development.
Disease Modeling: Given that DNA methylation has been implicated in various diseases, DNMT1 knockout 293T cells can be used to model certain pathological conditions. This model enables researchers to explore the contribution of methylation changes to disease phenotypes and develop strategies for therapeutic intervention.
For research use only. Not intended for any clinical use.
