This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 lung adenocarcinoma cell line, with targeted disruption of the JAG2 gene. The JAG2 Knockout A-549 Polyclonal Cells provide a loss-of-function model to investigate the biological roles of the JAG2-encoded Notch ligand in a lung cancer context. The polyclonal nature of the knockout population preserves cellular heterogeneity, making it suitable for population-level studies of pathway activity, phenotypic screening, and functional genomics experiments that require representation of diverse genetic backgrounds.
The host A-549 cell line was originally established from a pulmonary adenocarcinoma of a 58-year-old male and is widely employed as a model system for non-small cell lung cancer (NSCLC) research, epithelial cell biology, and drug sensitivity testing. A-549 cells retain key features of alveolar type II pneumocytes and exhibit characteristic epithelial morphology. They are extensively utilized to study oncogenic signaling, apoptosis, migration, and responses to chemotherapeutic agents, providing a physiologically relevant platform for investigating tumorigenic mechanisms and therapeutic vulnerabilities.
JAG2 encodes Jagged-2, a type I transmembrane ligand that mediates cell?Ccell communication via the Notch signaling pathway. Ligand engagement with NOTCH1 or NOTCH3 receptors on adjacent cells initiates sequential proteolytic cleavages: ectodomain shedding by ADAM10/ADAM17 metalloproteases, followed by intramembrane cleavage by the ??-secretase complex (including PSEN1). This releases the Notch intracellular domain (NICD), which translocates to the nucleus, forms a transcriptional activation complex with the DNA-binding factor RBPJ and coactivators such as MAML1, and drives expression of downstream targets including HES1, HEY1, MYC, CCND1, and SNAI1. JAG2 activity is modulated by upstream signals from TGFB1, EGF, and other Notch ligands, and is further regulated by glycosyltransferases LFNG and MFNG that modify receptor sensitivity. Knockout of JAG2 disrupts this signaling axis and may alter the balance of Notch-dependent transcriptional programs.
In A-549 lung adenocarcinoma cells, JAG2-mediated Notch activation has been implicated in regulating proliferation, apoptosis, epithelial?Cmesenchymal transition (EMT), and cancer stem cell maintenance. Abrogation of JAG2 function using this polyclonal knockout cell population provides a means to dissect the contribution of JAG2-specific Notch signaling to these processes in a lung cancer context. The model is therefore valuable for investigating the roles of JAG2 in tumorigenesis, metastatic potential, drug resistance, and the crosstalk with TGF-?? and other pathways that influence cellular plasticity and malignancy. It also offers a tool for comparative studies with other Notch ligands and receptors in adenocarcinoma.
Researchers can employ these cells in a broad range of functional assays, including Western blotting for Notch pathway components, RT-qPCR analysis of HES1 and HEY1 transcript levels, Notch reporter luciferase assays, and co-culture Notch activation experiments. Additional applications encompass migration and invasion assays to assess EMT-driven motility, proliferation and apoptosis measurements, immunofluorescence detection of NICD subcellular localization, flow cytometric profiling of JAG2 surface expression, and transcriptomic analyses such as RNA-seq. These methodologies enable detailed interrogation of JAG2-dependent signaling dynamics and cellular phenotypes in NSCLC models. For further technical information, please contact Ascent Research.
