CBR3 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered for disruption of the human CBR3 gene in the HAP1 near-haploid human cell line. This loss-of-function model enables researchers to study the biological consequences of CBR3 deficiency in a near-haploid genetic background that simplifies functional genomics approaches.
HAP1 cells are derived from the KBM-7 chronic myeloid leukemia line and exhibit a near-haploid karyotype with an adherent growth pattern. This unique genomic architecture facilitates gene disruption and allows the generation of polyclonal knockout populations with a high probability of functional gene inactivation across the cell pool, making HAP1 an ideal host for CRISPR/Cas9 knockout studies.
CBR3 encodes an NADPH-dependent carbonyl reductase that catalyzes the reduction of endogenous prostaglandins, such as prostaglandin E2, and xenobiotic substrates including the chemotherapeutic agent doxorubicin. The enzyme??s activity is transcriptionally regulated by NRF2 and the aryl hydrocarbon receptor in response to oxidative stress and electrophilic compounds. Downstream, CBR3 contributes to the inactivation of xenobiotics and reduction of reactive oxygen species levels. The protein functionally interacts with NADPH cofactor and cooperates with other carbonyl reductases like CBR1 and AKR1C3 within the arachidonic acid metabolism and xenobiotic detoxification pathways, intersecting with components such as PTGS2, PTGES, HPGD, CYP450s, NQO1, and GSTs.
In the HAP1 context, CBR3 knockout provides a powerful system to dissect the roles of this reductase in drug metabolism and prostaglandin biology. Given the near-haploid background, the polyclonal knockout population is well-suited for high-throughput screens aimed at identifying modifiers of doxorubicin sensitivity and regulators of oxidative stress. This model can help elucidate mechanisms underlying doxorubicin resistance in cancer, particularly in breast cancer where CBR3 expression influences prognosis and treatment response.
Researchers can employ CBR3 Knockout HAP1 Polyclonal Cells in a variety of functional assays, including western blotting and RT-qPCR for CBR3 expression analysis, prostaglandin reductase activity measurements, doxorubicin sensitivity assays for IC50 determination, and ROS level quantification using DCFDA. These cells are also compatible with pooled CRISPR screens followed by deep sequencing to identify genes that modulate CBR3-dependent cellular responses. Typical applications encompass functional studies of prostaglandin metabolism, investigation of chemotherapy resistance mechanisms, cancer biology research, and metabolic disorder modeling. For further information, please contact Ascent Research.
