The ANXA5 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered for loss-of-function studies of the ANXA5 gene, which encodes annexin A5. This product is generated by disrupting the target gene in the HEK293T host cell line, yielding a heterogeneous pool of edited cells suitable for investigating annexin A5-dependent processes without clonal selection bias. As polyclonal knockout cells, they provide a robust model for population-level analyses of gene disruption effects, enabling researchers to study pathways where ANXA5 plays a regulatory role.
The host cell line, HEK293T, is a widely used human embryonic kidney epithelial derivative expressing a mutant SV40 large T antigen. This modification permits episomal replication of vectors containing the SV40 origin, conferring high transfection efficiency and making these cells ideal for transient protein expression and retroviral production. HEK293T cells offer a versatile platform for analyzing gene function in a context that supports rapid experimental workflows, with well-characterized growth properties and compatibility with diverse molecular biology techniques.
Annexin A5 functions as a calcium-dependent phospholipid-binding protein that preferentially interacts with phosphatidylserine. Mechanistically, it binds to phosphatidylserine exposed on apoptotic cell surfaces in a Ca2?-regulated manner, thereby inhibiting blood coagulation by occupying sites required for assembly of coagulation factors such as Factor Xa and Factor Va. This interaction also regulates phagocytic clearance of apoptotic cells. Annexin A5 activity is modulated by upstream signals including Ca2? ions, EGF, TNF-??, and TGF-??1, and it acts downstream to suppress apoptosis, inhibit phospholipase A2 and protein kinase C, and interact with binding partners like actin, integrins, ANXA2, and S100A10.
Disruption of ANXA5 in HEK293T cells creates a system to dissect the role of annexin A5 in cellular homeostasis and disease-relevant pathways. Given the model??s background in a transformed epithelial line with high transfection capacity, the knockout cells enable dissection of annexin A5??s contributions to membrane repair, endocytosis, and phosphatidylserine signaling without interference from endogenous protein. The absence of annexin A5 may alter sensitivity to apoptotic stimuli or affect coagulation-related signaling, providing insights into disorders such as antiphospholipid syndrome, preeclampsia, systemic lupus erythematosus, and cancer, where dysregulated phosphatidylserine exposure and annexin function are implicated.
These polyclonal knockout cells are particularly suited for apoptosis detection using fluorescent annexin A5 in flow cytometry, calcium-dependent phospholipid binding assays, and coagulation assays to interrogate anticoagulation mechanisms. They also support immunofluorescence studies of membrane asymmetry and cell viability assessments in drug delivery research targeting phosphatidylserine. For further details on employing this model in your specific experimental context, please contact Ascent Research.
