This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human cell line, designed to disrupt the AKR1B10 gene. AKR1B10 encodes an aldo-keto reductase that functions as an NADPH-dependent retinaldehyde reductase and detoxification enzyme for reactive carbonyl species. Polyclonal knockout populations are generated by introducing CRISPR/Cas9 ribonucleoprotein complexes to induce targeted gene disruption, resulting in a heterogeneous pool of edited alleles. These cells serve as a loss-of-function model to investigate AKR1B10??s contributions to retinoic acid metabolism, aldehyde detoxification, and associated signaling networks. The population format supports robust functional genomics without requiring clonal isolation, making it suitable for pooled screening applications and downstream molecular assays.
The HAP1 host cell line is a human male chronic myelogenous leukemia (CML)-derived adherent line with a near-haploid karyotype. This genetic simplicity reduces functional redundancy and facilitates CRISPR-based phenotypic screens, as a single allelic disruption often yields a complete loss-of-function phenotype. HAP1 cells are widely employed in knockout validation, drug target discovery, and functional annotation of cancer-relevant genes. Their growth characteristics and stable karyotype enable reproducible experimental outcomes across multiple assay formats, including high-throughput imaging and biochemical analyses. The haploid background ensures that the AKR1B10 knockout population reliably reflects the consequences of gene disruption without the masking effects of a second allele.
AKR1B10 catalyzes the NADPH-dependent reduction of retinaldehyde to retinol, thereby limiting the pool of retinaldehyde available for oxidation to retinoic acid, the endogenous ligand for retinoic acid receptors (RARs) and retinoid X receptors (RXRs). This activity suppresses RAR/RXR-dependent transcription, influencing expression of target genes such as RAR?? and CYP26A1. In parallel, AKR1B10 detoxifies lipid peroxidation-derived aldehydes, including 4-hydroxynonenal (4-HNE), which attenuates oxidative stress and promotes cell survival. AKR1B10 is activated by transcription factors NRF2 and AP-1, and its expression is modulated by STAT3, PPAR??, and PPAR?? in response to electrophilic stress. Downstream, AKR1B10-mediated reduction of reactive carbonyls activates PI3K/AKT signaling and NF-??B, leading to upregulation of Cyclin D1, Bcl-2, and matrix metalloproteinases MMP-2 and MMP-9, which collectively drive proliferation, anti-apoptotic responses, and invasion.
In HAP1 cells, AKR1B10 disruption provides a critical tool for dissecting its role in cancers where it is frequently overexpressed, such as hepatocellular carcinoma, non-small cell lung cancer, breast cancer, and smoker??s lung cancer. By ablating AKR1B10, researchers can assess its impact on retinoic acid signaling through reporter assays and RT-qPCR for downstream targets like RAR??, evaluate changes in cellular sensitivity to oxidative stress, and measure effects on proliferation (MTS, BrdU), migration, and apoptosis (Annexin V, caspase assays). The haploid background enables clear interpretation of drug sensitivity and resistance phenotypes, facilitating the identification of synthetic lethal interactions or chemosensitization strategies. This model also supports validation of AKR1B10 as a therapeutic target and screening for small-molecule inhibitors.
The AKR1B10 Knockout HAP1 Polyclonal Cells are applicable across a wide range of experimental workflows. Typical applications include functional characterization of AKR1B10 in cancer cell biology using proliferation, migration, and invasion assays; investigation of retinoic acid metabolism by measuring aldo-keto reductase activity or retinoic acid-responsive luciferase reporters; and assessment of drug sensitivity in the presence of chemotherapeutic agents or targeted inhibitors. The cells can be employed in immunoblotting to confirm loss of AKR1B10 protein and to monitor downstream effectors such as Cyclin D1, Bcl-2, and MMP-2/MMP-9. For inquiries regarding technical specifications, customization, or bulk orders, please contact Ascent Research.
