Cat.No. : CSC-SC008942-1 Host Cell: HEK293
| Cat. No. | CSC-SC008942-1 |
| Description | HEK293-LRRK2 cell line is engineered to stably express human LRRK2. |
| Gene | LRRK2 |
| Gene Species | Homo sapiens (Human) |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| Shipping | Dry ice |
| 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. |
| 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. |
| 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 |
Leucine-rich repeat kinase 2 (LRRK2) is a crucial protein linked to the pathogenesis of Parkinson's disease (PD), with roles in dopaminergic neuronal vulnerability, oxidative stress (OS), mitochondrial dysfunction, and apoptosis. The researchers used CRISPR/Cas9 technology to generate LRRK2 knockout (KO) HEK-293 cells to study the cellular response to rotenone (ROT), a well-known inducer of OS and cell death. In their study, LRRK2 KO cells showed resistance to ROT-induced oxidative stress, mitochondrial damage, and apoptosis, compared to wild-type (WT) HEK-293 cells. This resistance was evidenced by unaltered OS markers, lack of mitochondrial dysfunction, and unchanged apoptosis-related pathways in the KO cells. The data further support the role of LRRK2 as a pro-apoptotic kinase, contributing to mitochondrial dysfunction and cell death in PD under stress conditions.
Figure 1. The researchers employed flow cytometry and fluorescence microscopy to assess the phosphorylation of LRRK2 at residue S935 in both LRRK2 WT and LRRK2 KO HEK-293 cells. They exposed these cells to rotenone (ROT) and found that LRRK2 KO cells showed no significant phosphorylation of S935-LRRK2, suggesting altered LRRK2 activation in the absence of the kinase. (Quintero-Espinosa DA, et al., 2023)
Creative Biogene offers a Human LRRK2 Stable Cell Line - HEK293, which can be used in similar studies exploring the role of LRRK2 in neurodegenerative diseases like Parkinson's. This cell line enables researchers to investigate LRRK2's involvement in oxidative stress and mitochondrial damage, contributing to a deeper understanding of the molecular mechanisms behind PD.
A: The stable expression of the LRRK2 gene in the Human LRRK2 Stable Cell Line-HEK293 is achieved by introducing the LRRK2 gene into HEK293 cells using specific vector systems, such as plasmids or viral vectors, to ensure long-term stable expression. This allows the cell line to continuously produce LRRK2 protein, providing a reliable model for studying its role in neurodegenerative diseases.
A: The application of the Human LRRK2 Stable Cell Line-HEK293 in Parkinson's disease research primarily involves modeling how LRRK2 gene mutations affect cellular functions, particularly in neurons. This cell line helps scientists understand the role of LRRK2 in the pathogenesis of Parkinson's disease, providing a foundation for developing therapies targeting this gene mutation.
A: The Human LRRK2 Stable Cell Line-HEK293 evaluates the activity of potential LRRK2 inhibitors by monitoring changes in LRRK2 protein activity. Researchers can use this cell line for high-throughput screening to identify compounds that inhibit LRRK2 activity and potentially slow or halt the progression of Parkinson's disease.
A: The Human LRRK2 Stable Cell Line-HEK293 contributes to studying the relationship between LRRK2 and apoptosis by providing a cellular model with stable LRRK2 expression. This helps reveal the role of LRRK2 in regulating the balance between cell death and survival, which is crucial for understanding apoptotic mechanisms in neurodegenerative diseases.
A: The Human LRRK2 Stable Cell Line-HEK293 can simulate LRRK2 activity under neuroinflammatory conditions, which is crucial for studying its role in neuroinflammatory responses. This cell line helps understand how LRRK2 participates in inflammatory signal transduction and its potential therapeutic value in neurodegenerative diseases.
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The Human LRRK2 Stable Cell Line-HEK293 provides a stable cellular model for studying the role of the LRRK2 gene in neurodegenerative diseases.
Contributes to a deeper understanding of the function of the LRRK2 gene in cellular physiology.
Using this cell line allows for the exploration of how LRRK2 mutations affect cell behavior and fate.
Provides a foundation for studying genetic variations of the LRRK2 gene and their impact on diseases.
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