The CBR4 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the human CBR4 gene. This polyclonal product is derived from the HAP1 cell line and contains a heterogeneous mixture of gene-disrupted cells, providing a robust model system without clonal selection. The knockout cell population is generated using CRISPR/Cas9-mediated gene disruption, enabling researchers to investigate the functional consequences of CBR4 deficiency across a bulk cell population.
The HAP1 host cell line is a near-haploid, fibroblast-like human cell line originally derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype simplifies gene editing and genetic screening, as only one allele requires modification to produce a knockout phenotype. HAP1 cells are widely employed in functional genomics studies, offering a streamlined platform for creating and analyzing knockout models. They retain key metabolic and signaling pathways relevant to human biology, making them suitable for studying a broad range of disease-associated genes.
CBR4 encodes a carbonyl reductase 4 enzyme that catalyzes the NADPH-dependent reduction of carbonyl substrates, including quinones, prostaglandins, and other endobiotic and xenobiotic compounds. This activity is central to cellular detoxification and lipid metabolism pathways. CBR4 expression is regulated by transcription factors such as NRF2, AhR, and PPAR??, which coordinate responses to oxidative stress and xenobiotics. Within the cell, CBR4 functions in conjunction with other oxidoreductases and detoxification enzymes, including CYP450 enzymes, glutathione S-transferases (GSTs), and NQO1, and it requires NADPH as a cofactor for its catalytic activity. Loss of CBR4 disrupts the conversion of cytotoxic carbonyls, leading to accumulation of reactive species and oxidative damage, which in turn can influence cell survival and drug sensitivity.
In the HAP1 cellular context, knockout of CBR4 provides a powerful tool to dissect mechanisms of carbonyl toxicity and oxidative stress response. The near-haploid background ensures a clean loss-of-function phenotype and facilitates subsequent genetic secondary screens or complementation studies. This model is particularly valuable for exploring how impaired carbonyl reduction contributes to pathophysiological states such as cancer progression, acquired drug resistance, and metabolic syndrome. By studying these polyclonal knockout cells, researchers can assess the impact of CBR4 deficiency on cellular fitness, metabolic reprogramming, and stress adaptation without the confounding effects of clonal variation.
Experimental applications of the CBR4 Knockout HAP1 Polyclonal Cells are extensive and include assays for carbonyl metabolism, drug metabolism, and oxidative stress biology. Typical assay formats include western blotting to confirm CBR4 protein loss, enzyme activity assays to quantify carbonyl reductase function, viability assays under carbonyl stress (e.g., menadione or 4-hydroxynonenal challenge), RT-qPCR for gene expression analysis of downstream targets, and metabolomics or flow cytometry to measure oxidative stress markers such as reactive oxygen species. These cells are also suitable for high-throughput genetic screens to identify modifiers of carbonyl sensitivity. For further technical information and ordering, please contact Ascent Research.
