GSTA2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line, designed for loss-of-function studies of the GSTA2 gene. This heterogeneous cell pool carries diverse CRISPR-mediated gene disruptions at the GSTA2 locus, offering a versatile model for functional genomics without single-cell cloning.
The HAP1 cell line is a near-haploid human chronic myelogenous leukemia line originally from a male patient. Its near-haploid karyotype facilitates gene knockout studies by reducing gene-copy complexity, enabling clear genotype-phenotype correlations. Widely used in cancer and signal transduction research, HAP1 cells provide a robust platform for high-throughput screening and editing.
GSTA2 encodes glutathione S-transferase alpha 2, a phase II detoxification enzyme that catalyzes the conjugation of reduced glutathione (GSH) to electrophilic xenobiotics and endogenous reactive oxygen species (ROS) products, including lipid peroxidation-derived aldehydes such as 4-hydroxynonenal (4-HNE), thereby facilitating their cellular excretion. GSTA2 transcription is tightly controlled by the KEAP1-NFE2L2 (NRF2)-antioxidant response element (ARE) signaling axis: upon oxidative or electrophilic challenge, NFE2L2 dissociates from KEAP1, translocates to the nucleus, dimerizes with small MAF transcription factors, and binds ARE sequences to induce GSTA2. Additional regulatory inputs from AHR, CAR, and PXR connect GSTA2 expression to xenobiotic metabolism pathways. Beyond its enzymatic function, GSTA2 physically interacts with JNK1 (MAPK8) and cooperates with GSTP1, modulating stress kinase signaling and overall detoxification network dynamics.
In the HAP1 background, GSTA2 knockout provides a definitive system to investigate the contribution of glutathione conjugation to drug resistance and redox homeostasis. Elevated GSTA2 expression is implicated in resistance to platinum-based chemotherapeutics (e.g., cisplatin) and anthracyclines (e.g., doxorubicin) in cancers such as hepatocellular carcinoma, lung adenocarcinoma, and ovarian carcinoma. The near-haploid HAP1 line eliminates confounding alleles, enabling direct correlation of GSTA2 loss with phenotypes in viability, intracellular ROS accumulation, and electrophile sensitivity assays. Thus, this model is highly suited for mechanistic studies of detoxification pathways and for identifying agents that circumvent GSTA2-mediated chemoresistance.
Applications include Western blotting and RT-qPCR for GSTA2 expression, CDNB-based enzymatic activity assays, and cell viability assays with cisplatin or doxorubicin. Additional readouts encompass NRF2/ARE luciferase reporter assays, ROS levels by flow cytometry, and JNK1 co-immunoprecipitation. These approaches support high-throughput screening for modulators of glutathione metabolism and chemoresistance. For product inquiries, please contact Ascent Research.
