The ANXA6 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disruption of the annexin A6 (ANXA6) gene, providing a loss-of-function model for studying ANXA6-dependent processes. This heterogeneous pool of gene-edited HEK293T cells avoids clonal selection artifacts and enables robust investigation of signal transduction, membrane trafficking, and disease biology.
HEK293T cells are human embryonic kidney cells transformed with adenovirus 5 DNA and expressing SV40 large T antigen. This line is widely used for virus production, recombinant protein expression, and cellular engineering due to its high transfectability and episomal plasmid replication. The SV40 large T antigen permits high-level protein expression and efficient generation of lentiviral and retroviral particles.
ANXA6 encodes a calcium-dependent membrane-binding scaffold that regulates endocytosis, exocytosis, and signal transduction. It negatively regulates EGFR signaling by promoting receptor internalization and degradation, thus dampening Ras-MAPK pathway activation. Activated by calcium influx, PKC, and EGFR ligands, ANXA6 interacts with p120 RasGAP, F-actin, S100A10, S100A11, and membrane phospholipids to organize signaling complexes and terminate EGFR-driven signals. ANXA6 knockout is expected to impair EGFR downregulation, causing sustained Ras-Raf-MEK-ERK signaling and altered actin cytoskeleton dynamics.
In HEK293T cells, loss of ANXA6 disrupts negative regulation of EGFR signaling, leading to elevated Ras-MAPK activity that can influence proliferation, survival, and viral production efficiency. The polyclonal knockout format provides internal control against clonal variation, ensuring phenotype robustness. This model is particularly suited for studying receptor trafficking, signal termination, and the interplay between membrane dynamics and signaling in an epithelial context.
The ANXA6 Knockout HEK293T Polyclonal Cells enable applications such as EGFR regulatory studies, Ras-MAPK signaling analysis, membrane trafficking and endocytosis research, and cholesterol homeostasis investigation. Representative assays include Western blotting for EGFR and phospho-ERK, RT-qPCR of MAPK targets, immunofluorescence for EGFR localization, co-immunoprecipitation with p120 RasGAP or S100 proteins, flow cytometry for surface EGFR, phospho-signaling analysis, and apoptosis assays. These cells also support cancer and neurodegeneration research and drug screening. For further details, contact Ascent Research.
