The ATP11A Knockout A-549 Polyclonal Cells constitute a genetically engineered cell population derived from the human A-549 lung adenocarcinoma epithelial cell line, featuring CRISPR/Cas9-mediated disruption of the ATP11A gene. This polyclonal knockout model is produced by introducing targeted gene edits via CRISPR/Cas9 ribonucleoprotein delivery, followed by selection to enrich for cells with loss-of-function mutations in ATP11A. As a heterogeneous pool, the polyclonal format preserves the biological variability inherent to a population of knockout cells, making it suitable for studying gene function without clonal artifacts. The disruption of ATP11A eliminates the expression of functional flippase, thereby abolishing the active transport of phosphatidylserine from the outer to the inner leaflet of the plasma membrane. This model provides a robust platform for investigating the roles of phospholipid asymmetry in cellular signaling and disease processes.
The host A-549 cell line was originally established from the lung adenocarcinoma tissue of a 58-year-old Caucasian male and serves as a widely used in vitro model for non-small cell lung cancer (NSCLC). These adherent epithelial cells retain key properties of alveolar type II pneumocytes, including the ability to form confluent monolayers and express certain surfactant proteins. A-549 cells are commonly employed in cancer biology research to dissect oncogenic signaling pathways, drug responses, and metastasis mechanisms. Their genetic background harbors mutations typical of lung adenocarcinoma, such as a KRAS G12S mutation, which makes them particularly relevant for studying tumorigenesis driven by RAS pathway activation. The integration of the ATP11A knockout into this established lung cancer model enables the elucidation of flippase-dependent functions within a tumor-relevant cellular environment.
ATP11A encodes a P4-type ATPase that functions as a phosphatidylserine flippase, actively translocating phosphatidylserine from the exoplasmic to the cytoplasmic leaflet of the plasma membrane. This transport is strictly dependent on the cofactor CDC50A, a member of the TMEM30 family, which is required for proper folding, trafficking, and ATPase activity of ATP11A. Under physiological conditions, ATP11A in complex with CDC50A maintains the asymmetric distribution of phosphatidylserine, confining this phospholipid to the inner leaflet. In contrast, scramblases such as TMEM16F can non-selectively equilibrate phospholipids across the bilayer upon activation. ATP11A opposes scramblase activity, thereby preventing the constitutive externalization of phosphatidylserine. The functional interplay between ATP11A, CDC50A, and scramblases represents a core network governing membrane lipid dynamics, with direct implications for apoptosis signaling and intercellular communication.
The loss of ATP11A flippase activity in A-549 cells leads to the sustained exposure of phosphatidylserine on the cell surface, a hallmark of apoptotic cells. In the context of lung adenocarcinoma, this aberrant lipid distribution can modulate tumor-host interactions, influencing immune recognition, phagocytosis by macrophages, and the coagulation cascade. Moreover, phosphatidylserine exposure has been linked to enhanced invasiveness and drug resistance in cancer cells, potentially through the activation of signaling pathways such as PI3K/AKT. Research on ATP11A-related neurodevelopmental disorders and congenital hemolytic anemia further underscores its critical role in cellular homeostasis. By employing this knockout model, scientists can dissect how ATP11A deficiency impacts tumor cell behavior, apoptotic clearance, and the response to chemotherapeutic agents in a lung adenocarcinoma background.
This polyclonal knockout cell product is ideally suited for a broad range of functional studies employing assays such as annexin V binding and flow cytometry to quantify phosphatidylserine externalization, as well as western blotting and RT-qPCR to validate gene disruption at the protein and mRNA levels. Immunofluorescence microscopy enables visualization of ATP11A localization and membrane asymmetry changes, while apoptosis assays and lipidomic profiling provide deeper insights into cell death mechanisms and global lipid remodeling. Researchers can utilize these cells to investigate the crosstalk between flippase activity and oncogenic signaling, to screen for modulators of phosphatidylserine-dependent drug resistance, or to evaluate the impact of ATP11A knockout in co-culture systems with immune cells. For additional technical specifications or custom requests, please contact Ascent Research.
