The CBR1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the human CBR1 gene. This product provides a mixed pool of HAP1 cells carrying heterogeneous loss-of-function mutations introduced by CRISPR/Cas9, enabling robust functional studies without clonal isolation. The polyclonal format preserves genetic diversity while minimizing clonal artifacts, making it suitable for pooled genetic screens and population-level analyses of gene function.
The HAP1 cell line is a near-haploid, fibroblast-like model derived from the male chronic myelogenous leukemia cell line KBM-7. It harbors a mutated p53 tumor suppressor, which facilitates genetic manipulability and tolerance to chromosomal alterations. HAP1 cells are extensively employed in functional genomics, haploid genetic screens, and drug?Cgene interaction studies due to their stable near-haploid karyotype and rapid proliferation.
CBR1 encodes an NADPH-dependent carbonyl reductase that catalyzes the reduction of a broad range of endogenous and exogenous carbonyl substrates. A key function is the conversion of prostaglandin E2 (PGE2) to prostaglandin F2?? (PGF2??), thereby modulating inflammatory signaling. The enzyme also detoxifies reactive aldehydes such as 4-hydroxynonenal (4-HNE) generated during lipid peroxidation and metabolizes anthracycline chemotherapeutics like doxorubicin to the less potent doxorubicinol. CBR1 expression is transcriptionally regulated by the NRF2 transcription factor in response to oxidative stress, placing it downstream of the KEAP1-NRF2 antioxidant pathway. Its activity depends on the NADPH cofactor and is linked to upstream prostaglandin synthesis by COX-1 and COX-2, as well as PGE2 synthase. Downstream effectors include the prostaglandin transporter-mediated export of PGF2?? and detoxified lipid peroxidation products.
In the HAP1 leukemic background, disruption of CBR1 is expected to alter prostaglandin metabolism, shifting the PGE2/PGF2?? balance and potentially affecting inflammatory responses and cell survival pathways. The p53-deficient context may further modulate the impact of carbonyl stress, making this knockout model particularly relevant for studying oxidative stress response and drug sensitivity. Given the role of CBR1 in doxorubicin metabolism, HAP1 CBR1 knockout cells provide a valuable system to investigate mechanisms of anthracycline-induced cardiotoxicity and resistance.
This polyclonal knockout population is well-suited for a variety of functional assays, including Western blotting and RT-qPCR to confirm CBR1 disruption, ELISA-based quantification of PGE2 and PGF2?? to assess prostaglandin metabolism, and cell viability assays under oxidative stress (e.g., H2O2) or chemotherapeutic challenge (e.g., doxorubicin cytotoxicity measured by MTT). Additionally, ROS detection via DCFDA and flow cytometric apoptosis analysis (Annexin V/PI) can be employed to dissect pathways of cell death. These tools make the cells invaluable for mechanistic studies in inflammation, cancer biology, and drug toxicity. For further details, please contact Ascent Research.
