The BZW2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the BZW2 gene to generate a loss-of-function model. This product consists of a heterogeneous pool of edited cells, enabling study of BZW2-dependent signaling without clonal isolation artifacts. The polyclonal format preserves population-level diversity while eliminating BZW2 protein expression, suitable for pooled functional studies and pathway-level analyses.
HEK293T cells, derived from human embryonic kidney cells, stably express the SV40 large T antigen, which supports episomal replication of plasmids containing the SV40 origin for high transgene expression and efficient viral production. These adherent epithelial cells are highly transfectable and widely used for protein expression, viral packaging, and cell biology applications. Their robust translational machinery and well-mapped signaling networks make them an excellent host for studying amino acid sensing and the integrated stress response.
BZW2 acts as a competitive inhibitor of GCN2 kinase (EIF2AK4), a primary sensor of amino acid deprivation. Under nutrient-sufficient conditions, BZW2 binds GCN2 and suppresses its activity, thereby limiting phosphorylation of eIF2?? and downstream ATF4-dependent transcription associated with the integrated stress response (ISR). Amino acid scarcity triggers BZW2 dissociation, activating GCN2, which phosphorylates eIF2?? and induces ATF4 and stress-responsive genes such as CHOP. BZW2 also interacts with eIF5 and the eIF2 complex to modulate translation initiation. This regulatory node is integrated with mTORC1 signaling, where mTORC1 influences BZW2 in response to amino acid and energy cues, shaping global protein synthesis.
In HEK293T cells, BZW2 knockout establishes constitutive, low-level GCN2 activation and ISR engagement under normal culture conditions, sensitizing cells to amino acid starvation and ER stress. This model allows dissection of BZW2??s buffering role on stress signaling in a cell type central to protein production. High transfectability enables complementation with wild-type or mutant BZW2 for structure-function studies and drug target validation. The polyclonal nature avoids clonal variation artifacts, ensuring phenotypes arise from BZW2 loss.
Researchers can employ these cells to study tumor suppressor mechanisms (relevant to colorectal cancer, hepatocellular carcinoma, and lung adenocarcinoma) and amino acid metabolism disorders. Typical assays include amino acid starvation experiments, polysome profiling to measure global translation shifts, and Western blotting/RT-qPCR for phospho-eIF2??, ATF4, and downstream targets. Co-immunoprecipitation confirms BZW2?CGCN2 interaction, while ATF4-luciferase reporters quantify ISR activity. Cell viability assays under ER stress assess functional consequences. For more information or to place an order, please contact Ascent Research.
