HMGCR Knockout 293T Cell Line, Heterozygous

Cell Lines & Derivatives

HMG-CoA reductase is the rate-limiting enzyme for cholesterol synthesis and is regulated via a negative feedback mechanism mediated by sterols and non-sterol metabolites derived from mevalonate, the product of the reaction catalyzed by reductase. Normally in mammalian cells this enzyme is suppressed by cholesterol derived from the internalization and degradation of low density lipoprotein (LDL) via the LDL receptor. Competitive inhibitors of the reductase induce the expression of LDL receptors in the liver, which in turn increases the catabolism of plasma LDL and lowers the plasma concentration of cholesterol, an important determinant of atherosclerosis. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.

1 vial parental cell line and 1 vial knockout cell line

Upon arrival, it should be maintained in DMEM medium with 10%(v/v) fetal bovine serum and 100U penicillin-streptomycin, at 37°C with 5% CO₂ condition.
1. Thaw the vial in 37°C water bath , and shake it to melt as soon as possible.
2. Transfer the cell suspension to a 15mL conical tube with pre-warmed 5mL complete me-
        dium and centrifuge 1000rpm for approximately 5 minutes at room temperature.
3. Remove and discard the supernatant.
4. Resuspend the cell pellet with 1mL pre-warmed complete medium and seed in 10cm dish.
5. Add 8-10mL of complete medium.
6. Incubate the culture at 37°C incubator with 5% CO₂.
7. A subcultivation ratio of 1:2-1:4 is recommended.