GSTM3 Knockout HEK293T Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population of the human embryonic kidney HEK293T cell line. The knockout targets the GSTM3 gene, which encodes glutathione S-transferase mu 3, a phase II detoxification enzyme. This polyclonal pool offers a loss-of-function model without single-cell cloning, suitable for population-level phenotypic studies. The knockout is generated via CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous mixture of cells with ablated GSTM3 expression.
HEK293T cells are an extensively characterized human embryonic kidney epithelial line stably expressing the SV40 large T antigen. This feature enhances episomal replication and protein expression, making these cells ideal for recombinant protein production, viral packaging, and functional genomics applications. The epithelial origin and robust growth of HEK293T provide a reproducible platform for investigating cellular detoxification and oxidative stress pathways. Moreover, they possess an intact KEAP1-Nrf2 axis that transcriptionally regulates GSTM3, thus offering a physiologically relevant context for knockout studies.
GSTM3 catalyzes the conjugation of reduced glutathione (GSH) to electrophilic substrates, including xenobiotics, carcinogens, and lipid peroxidation products, for detoxification and excretion. Its expression is driven by NFE2L2 (Nrf2) upon oxidative/electrophilic stress and is also regulated by AHR. GSTM3 interacts with GSTM1, GSTM2, and MAP3K5 (ASK1), linking it to stress kinase signaling. Through these activities, GSTM3 modulates drug metabolism (CYP2E1), limits ferroptosis by reducing lipid peroxides, and integrates into the Nrf2 antioxidant network involving GSH, GPX, and NQO1.
Disrupting GSTM3 in HEK293T cells ablates a critical detoxification node, sensitizing cells to oxidative and electrophilic insults. This model enables dissection of GSTM3-specific roles in the Nrf2 response, distinct from other GSTs. The population is ideal for studying ferroptosis, chemoresistance, and metabolic activation of xenobiotics, as HEK293T supports high-throughput screens. Additionally, GSTM3?CMAP3K5 interactions can be examined to reveal crosstalk between detoxification and apoptosis.
Typical applications include oxidative stress profiling, drug metabolism and toxicity screening, cancer biology (ferroptosis, chemoresistance), and Nrf2 pathway studies. Compatible assays: western blot, RT-qPCR, GST activity, viability under H2O2 or paraquat, ROS (DCFH-DA), GSH/GSSG ratio, and Nrf2 immunofluorescence. For further details, contact Ascent Research.
