This product is a CRISPR/Cas9-edited polyclonal ALPP knockout cell population derived from the human A-549 lung adenocarcinoma cell line. The cells harbor a targeted disruption of the ALPP gene, encoding placental alkaline phosphatase, generated via CRISPR/Cas9-mediated genome editing without clonal isolation. This heterogeneous loss-of-function pool enables analysis of ALPP-dependent phenotypes in an alveolar epithelial context.
The parental A-549 cell line originates from a lung adenocarcinoma of a 58-year-old Caucasian male and serves as a widely used in vitro model of human type II alveolar epithelium. It is extensively applied in respiratory disease and oncology research, particularly for studying lung adenocarcinoma progression, epithelial cell biology, and cancer-associated signaling.
ALPP encodes a GPI-anchored alkaline phosphatase localized to lipid rafts, where it homodimerizes and interacts with caveolin-1. The enzyme hydrolyzes phosphate monoesters and dephosphorylates extracellular substrates, including pyridoxal phosphate and phospho-??-catenin, thereby modulating phosphate metabolism and signal transduction. ALPP transcription is regulated by cAMP, steroid hormones, and the Sp1 factor. Its activity influences Wnt/??-catenin signaling by dephosphorylating phospho-??-catenin, which stabilizes ??-catenin, and BMP signaling through effects on pyridoxal phosphate utilization and downstream Smad1/5/8. Thus, ALPP knockout disrupts dephosphorylation, leading to accumulation of phosphorylated substrates and altered pathway dynamics.
In A-549 lung adenocarcinoma cells, ALPP knockout provides a system to investigate its contribution to tumorigenesis. Loss of enzymatic activity impairs pyridoxal phosphate dephosphorylation, reducing vitamin B6 cofactor availability, and increases phospho-??-catenin, potentially attenuating Wnt/??-catenin-driven transcription. These changes may impact cell proliferation, migration, and invasion, offering a relevant model for examining ALPP??s role in lung adenocarcinoma progression and its potential as a biomarker or drug target.
This polyclonal knockout cell population is suitable for diverse assays: alkaline phosphatase activity assays confirm functional knockout, while Western blotting, RT-qPCR, and RNA-seq probe gene expression changes. Phosphoproteomics and pyridoxal phosphate measurement assess substrate phosphorylation states. Functional assays, including cell proliferation, migration/invasion, and Wnt/BMP reporter assays, enable detailed phenotypic analysis. Applications span biomarker validation, drug discovery, and pathway modulation studies. For further information, please contact Ascent Research.
