The CBR4 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the CBR4 gene in the HeLa cell line. This polyclonal pool serves as a loss-of-function model to study the mitochondrial carbonyl reductase CBR4 without clonal isolation, preserving population-level heterogeneity for robust functional analyses.
HeLa cells, derived from HPV18-positive cervical adenocarcinoma, are an aneuploid epithelial cell line known for their vigorous growth and broad utility in cancer biology, cell signaling, and drug discovery. Their genetic tractability and extensive characterization make them a preferred host for generating knockout models to dissect metabolic and disease-relevant pathways.
CBR4 encodes a mitochondrial short-chain dehydrogenase/reductase that catalyzes the reduction of 3-ketoacyl-CoA intermediates during long-chain fatty acid beta-oxidation, a critical step for complete fatty acid degradation and mitochondrial ATP production. CBR4 expression is regulated by PPARG coactivator 1 alpha (PPARGC1A) and peroxisome proliferator-activated receptors (PPARs), and its activity is modulated by the cellular energy sensor AMPK. Within the mitochondrial matrix, CBR4 physically interacts with the mitochondrial trifunctional protein (MTP) and enoyl-CoA hydratase (ECHS1), and functions cooperatively with HADHA, HADHB, and ACADVL. Consequently, CRISPR/Cas9-mediated disruption of CBR4 impairs fatty acid oxidation, leading to altered cellular lipid profiles and diminished oxidative ATP synthesis.
In the HeLa cellular context, CBR4 knockout constitutes a powerful tool to investigate mitochondrial metabolic reprogramming in cancer. HeLa cells depend on both glycolysis and oxidative phosphorylation, and loss of CBR4 may induce compensatory shifts in substrate utilization, revealing metabolic vulnerabilities under nutrient or drug stress. This model therefore enables exploration of how mitochondrial dysfunction influences cancer cell proliferation, lipid homeostasis, and survival mechanisms.
Researchers can employ these polyclonal knockout cells across a spectrum of functional assays, including Western blotting and RT-qPCR for target validation, 14C-oleate-based fatty acid oxidation assays to quantify metabolic flux, and Seahorse mitochondrial stress tests to profile oxygen consumption. Complementary lipidomics analyses, ATP quantification, and cell viability studies under metabolic challenge provide comprehensive readouts. Moreover, the cells are well suited for screening small-molecule modulators of mitochondrial metabolism and for investigating xenobiotic detoxification pathways mediated by short-chain dehydrogenases/reductases. For additional product information, please contact Ascent Research.
