The CCDC47 Knockout A-549 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the CCDC47 gene in the A-549 human lung adenocarcinoma cell line. This pooled knockout format provides a heterogeneous cell population harboring diverse loss-of-function mutations at the target locus, enabling robust characterization of gene function without clonal selection artifacts. The polyclonal population is suitable for studying CCDC47-dependent phenotypes in a genetically diverse cellular context, reflecting the complexity of epithelial cancer models. The knockout model is generated using transient delivery of CRISPR/Cas9 ribonucleoprotein complexes, ensuring efficient gene disruption while maintaining host cell line characteristics.
The A-549 cell line is a widely used epithelial model derived from lung adenocarcinoma tissue of a 58-year-old male patient. These cells exhibit adherent growth and retain key features of type II alveolar epithelium, including lamellar body formation and surfactant production. A-549 cells serve as a foundational in vitro system for non-small cell lung cancer research, recapitulating oncogenic signaling networks, tumor-stromal interactions, and therapeutic responses relevant to clinical disease. Their well-characterized genetic background and extensive experimental history make them ideal for investigating gene function in lung adenocarcinoma pathogenesis.
CCDC47 encodes a coiled-coil domain-containing protein localized to the endoplasmic reticulum, where it plays indispensable roles in ER proteostasis and calcium homeostasis. CCDC47 functions as a cofactor for the ER chaperone HSPA5/BiP, facilitating proper protein folding and assembly. It also interacts with SERCA2 and calnexin to modulate ER calcium storage and release. Under ER stress conditions, upstream regulators ATF4, ATF6, and spliced XBP1 activate the unfolded protein response by transcriptionally upregulating CCDC47 along with canonical UPR effectors. Loss of CCDC47 disrupts calcium-dependent chaperone activity, leading to sustained activation of PERK, IRE1, and ATF6 signaling branches. This triggers downstream pro-apoptotic mediators CHOP/DDIT3, Caspase-12, and BAX/BCL2 imbalance, sensitizing cells to ER stress-induced death.
In the A-549 lung cancer context, CCDC47 knockout generates a constitutive ER stress phenotype that profoundly impairs tumor cell fitness. Disrupted calcium signaling and chronic UPR activation compromise cell proliferation, migration, and clonogenic survival, while elevating susceptibility to chemotherapeutic agents and targeted therapies. This model enables dissection of ER stress adaptive mechanisms exploited by non-small cell lung cancer cells for survival. Given CCDC47’s emerging links to hepatocellular carcinoma and ER stress-related disorders, the polyclonal knockout population provides a versatile platform for investigating shared stress response pathways across cancer types.
Researchers can employ this knockout model for a wide range of functional studies, including characterization of ER stress signaling dynamics via Western blotting for BiP and CHOP, calcium flux imaging with fluorescent indicators, and apoptosis quantification using Annexin V staining. Proliferation and migration deficits can be assessed by MTS and Transwell assays, respectively, while qPCR profiling of UPR target genes validates pathway engagement. Applications extend to drug resistance screens, synthetic lethal interaction mapping, and high-content functional genomics in lung cancer biology. For detailed technical specifications and ordering information, please contact Ascent Research.
