APP Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered from the CAL-27 human tongue squamous cell carcinoma cell line. The APP gene has been disrupted via CRISPR/Cas9 to generate a loss-of-function model for investigating the roles of amyloid precursor protein in cancer biology and signal transduction. This polyclonal population retains the genetic heterogeneity inherent to mixed edited pools, making it suitable for bulk functional assays without the selective pressure of clonal isolation.
The host cell line, CAL-27, is derived from a human tongue squamous cell carcinoma and carries a mutant TP53 gene, rendering it tumorigenic. CAL-27 cells are a well-established in vitro model for oral squamous cell carcinoma, characterized by aggressive growth, and are widely employed in studies of cell adhesion, migration, invasion, and signaling pathways relevant to head and neck cancers.
APP encodes a type I transmembrane protein that undergoes sequential cleavage by ??-, ??-, and ??-secretases, including BACE1, ADAM10, PSEN1, and NCSTN, yielding soluble ectodomains (sAPP??/??) and the AICD intracellular domain. AICD interacts with Fe65 and Tip60 to form a transcriptional complex that regulates genes controlling cell adhesion and proliferation. APP also engages integrin beta1 and LRP1, modulating integrin and PI3K/AKT signaling. Upstream regulators such as SP1, NF-kB, IL-6, and TNF-alpha influence APP expression, while downstream pathways involve EGFR, beta-catenin, GSK3B, AKT, and ERK1/2. Thus, APP sits at the nexus of multiple networks, including Notch, Wnt, and MAPK/ERK, affecting cell motility and survival.
In the context of CAL-27 oral cancer cells, APP is implicated in promoting cell adhesion, migration, and invasion, partly through PI3K/AKT and integrin signaling. Disrupting APP in this polyclonal knockout population enables researchers to dissect its contribution to the malignant phenotype, including its effects on the AICD-Fe65-Tip60 transcriptional complex and downstream effectors like EGFR and beta-catenin. This model offers a valuable tool to explore the crosstalk between neurobiological APP processing and cancer progression.
This product facilitates the study of APP function in oral cancer, with applications in cell adhesion, migration, and invasion assays. It supports drug testing of secretase inhibitors, analysis of APP processing via western blotting for full-length APP and fragments (CTF, sAPP), and examination of AICD-mediated transcription through co-immunoprecipitation of Fe65 complexes. Supplementary assays include RT-qPCR for APP, EGFR, and LRP1; immunofluorescence for integrins; proliferation assays; and phospho-AKT/ERK analysis. Gamma-secretase activity can be assessed using biochemical methods. For technical inquiries, please contact Ascent Research.
