Cas9 Stable Cell Line - C2C12

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.