Cas9 Stable Cell Line - C2C12
Cat.No. : CSC-RO0186 Host Cell: C2C12
Size: >1x10^6 cells/vial Validation: T7 Endonuclease I assay
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Cell Line Information
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Datasheet
| Cat. No. | CSC-RO0186 |
| Product Type | Cas9 overexpression stable cell line |
| Introduction | Clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 is a gene-editing technology that contains two essential components: a guide RNA (gRNA) to match a target gene, and the Cas9 (CRISPR-associated protein 9) endonuclease which causes a double-stranded DNA break, allowing modifications to the genome via nonhomologous end joining (NHEJ) or homology-directed repair (HDR). |
| Cell Line Information | C2C12-Cas9 cell line is engineered to stably overexpress Cas9 nuclease. The Cas9 nuclease in C2C12-Cas9 cell line has been functionally validated using T7 Endonuclease I assay. In combination with separately transfected sgRNAs, C2C12-Cas9 cell line can be used to efficiently generate targeted genomic modifications including gene knockout, gene knockin, gene mutagenesis, gene tagging etc. It is also an ideal cell line model for sgRNA screening and validation, either individually or in pools. |
| Target Gene | Cas9 |
| Host Cell | C2C12 |
| Applications | 1) CRISPR genome editing, such as gene knockout (KO), gene knockin (KI), gene mutagenesis, gene tagging etc. 2) High-throughput sgRNA screening and validation |
| Quality Control | 1) T7E1 assay 2) Mycoplasma detection |
| Size Form | One vial of frozen cells, typically >1x10^6 cells/vial |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| 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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Case Study
Applications
The division and proliferation of skeletal muscle cells are strictly controlled. The function of SET domain containing 2 (Setd2) in myoblast development and proliferation was examined by the researchers. They observed abnormalities in myotube formation and downregulation of myosin heavy chain (MHC) and Myogenin (MyoG) expression after silencing Setd2 in C2C12 skeletal muscle cells using CRISPR/CAS9. Reduced proliferation, decreased histone 3 phosphorylation, impaired G1/S- and G2/M-phase transitions, and decreased production of cyclin D1, CDK4, CDK6, and cyclin E2 were the results of silencing setd2. Moreover, there was an upregulation of p21, which signifies cell cycle arrest. The work emphasizes how important Setd2 is for myoblast development and proliferation.
Figure 1. The researchers set out to generate Setd2−/− C2C12 cell lines. They transfected C2C12 cells with plasmids encoding Setd2 sgRNA and Cas9 in order to employ CRISPR/Cas9 genome editing. Sorted, isolated, and cultivated cells were found to be GFP-positive. Using PCR, sequencing, and Intelligent software, mutations were verified. By using western blot, clones with frame-shift mutations were confirmed to be Setd2−/−. After being examined, no off-target effects were discovered at the top possible locations. (Yi X, et al., 2017)
Try Creative Biogene's Cas9 stable cell line - C2C12 cell line! This cell line has stably expressed Cas9 protein, eliminating the steps of transfecting Cas9 protein or constructing Cas9 expression vectors, saving time and effort. With this cell line, you can directly perform gene editing experiments, reducing the possibility of additional steps and variations. This will improve the efficiency and consistency of the experiment, allowing you to focus more on data analysis and interpretation, thereby accelerating your research progress.
1. CRISPR-Cas9 Gene Editing: Targeted genome editing is achieved through the use of CRISPR-Cas9 technology in the C2C12 stable cell line.
2. Cas9 Endonuclease Expression: Cas9 endonuclease expression is achieved through genetic engineering of C2C12 cells, which allows for accurate DNA cleavage.
3. Research on Muscle Development: C2C12 cells are frequently employed in investigations on the growth and regeneration of muscles.
4. Mouse Myoblast Cell Line: C2C12 cells, which are derived from mouse skeletal muscle satellite cells, can develop into myotubes in vitro.
5. RNA-Guided DNA Cleavage: Cas9 in C2C12 cells cleaves particular DNA sequences under the supervision of RNA, allowing for precise genetic alterations.
For research use only. Not intended for any clinical use.
