The ANXA11 knockout HEK293T polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the target gene ANXA11, creating a loss-of-function model in a widely used human cell background. This polyclonal knockout product provides a heterogeneous pool of edited cells, enabling the study of ANXA11-dependent processes without clonal selection bias. The CRISPR/Cas9-mediated gene disruption in these polyclonal cells facilitates investigation of ANXA11’s role in calcium-regulated membrane events and disease-relevant pathways.
The host cell line, HEK293T, is a human embryonic kidney epithelial cell derivative transformed with adenovirus type 5 DNA. These cells constitutively express the SV40 large T antigen, which allows episomal replication of plasmids containing the SV40 origin of replication, thereby enhancing recombinant protein expression and enabling efficient lentivirus production. HEK293T cells are extensively characterized and serve as a robust platform for studying signal transduction, membrane dynamics, and protein interactions due to their ease of transfection and well-documented cellular machinery.
ANXA11 encodes a calcium-dependent phospholipid-binding protein that participates in multiple cellular processes including membrane trafficking, apoptosis, cell cycle regulation, and exosome biogenesis. It interacts with key molecular partners such as ALG2 (PDCD6), S100A6 (calcyclin), calcium ions, phospholipids, and F-actin, forming complexes that mediate membrane repair and vesicle organization. Upstream, ANXA11 is regulated by calcium influx, DNA damage signals, and mitogens. Downstream, it influences caspase-3 activation and autophagy marker LC3B turnover, integrating signals from the apoptosis and autophagy pathways. Mechanistically, ANXA11 acts as a scaffold linking calcium signaling to membrane dynamics and stress responses.
In the HEK293T cellular context, knockout of ANXA11 disrupts calcium-mediated membrane dynamics and may impair stress-induced apoptosis and autophagy, providing a powerful model to dissect its role in neurodegeneration. Given the association of ANXA11 mutations with amyotrophic lateral sclerosis (ALS) and frontotemporal dementia, this polyclonal knockout population allows researchers to examine how loss of ANXA11 function alters cellular resilience to proteotoxic stress, membrane repair efficiency, and intercellular communication via exosomes. The model is particularly valuable for studying protein aggregation phenotypes and calcium dysregulation without the confounding effects of clonal variation.
This product is ideally suited for a range of advanced research applications, including the investigation of ALS disease mechanisms, calcium-dependent membrane trafficking, apoptosis and autophagy signaling, and protein aggregation studies. Representative assays include Western blotting to confirm ANXA11 ablation, calcium influx measurements using fluorescent indicators, apoptosis assays via Annexin V/PI staining, autophagy flux analysis based on LC3B turnover, co-immunoprecipitation of ALG2 and S100A6 complexes, and immunofluorescence staining for vesicle markers to monitor trafficking defects. Drug screening for ALS therapeutics can be performed using this model to identify compounds that rescue ANXA11-related phenotypes. For further technical details, please contact Ascent Research.
