The ATP11B Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited population derived from A-549 lung adenocarcinoma cells, disrupting the ATP11B gene encoding a phospholipid flippase. This polyclonal knockout model abrogates ATP11B-mediated translocation of phosphatidylserine (PS), leading to constitutive PS externalization. Generated via transient Cas9/sgRNA delivery without clonal selection, the population retains genetic heterogeneity while providing robust target-gene disruption, suitable for functional studies in a lung cancer context.
The A-549 cell line, established from a 58-year-old Caucasian male with lung adenocarcinoma, serves as a classical model for human alveolar type II pneumocytes, exhibiting lamellar bodies and surfactant synthesis. Widely utilized in oncology, drug discovery, and respiratory biology, A-549 cells are adherent, amenable to genetic manipulation, and characterized extensively in chemotherapeutic response and signaling pathways.
ATP11B is a P4-ATPase flippase that partners with the obligatory ??-subunit CDC50A (TMEM30A) to catalyze ATP-dependent inward translocation of PS, preserving membrane lipid asymmetry. This activity is essential for vesicular trafficking, cell polarity, and the timely clearance of apoptotic cells. Loss of ATP11B causes PS to remain on the outer leaflet, engaging PS receptors such as TIM4 and modulating apoptotic signaling. ATP11B is inhibited by caspase cleavage during apoptosis and is regulated by protein kinase C. It operates alongside the scramblase ANO6 (TMEM16F), which randomizes phospholipid distribution, and interacts with CDC50A for proper folding and trafficking. These molecular interactions position ATP11B at the interface of membrane dynamics, cell survival, and immune recognition.
In the A-549 background, ATP11B knockout disrupts membrane asymmetry, offering a defined system to probe PS-dependent phenomena in lung adenocarcinoma. The model facilitates exploration of altered cell polarity, migration, and apoptotic resistance relevant to cancer progression. Constitutive PS exposure may influence immune evasion and drug sensitivity, providing insights into how lipid flippases contribute to tumor biology. This engineered cell population thus serves as a powerful tool for dissecting ATP11B??s roles in endocytosis, lipid signaling, and epithelial pathophysiology.
These polyclonal knockout cells are applicable to apoptosis assays (Annexin V staining, flow cytometry), flippase activity measurements, western blotting, and cell migration studies. They enable investigations into membrane asymmetry??s impact on drug resistance, PS-mediated signaling, and immune modulation. Researchers can interrogate ATP11B??s interaction with CDC50A and ANO6, and study its role in cancer cell biology, neurodevelopmental disorders, and lipid transport pathways. For technical specifications or ordering, please contact Ascent Research.
