The JAG2 Knockout MCF-7 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of human MCF-7 breast adenocarcinoma cells carrying a targeted disruption of the JAG2 gene. This loss-of-function model enables investigation of Notch signaling pathway dynamics by eliminating the expression of the JAG2-encoded Notch ligand. The polyclonal nature of the knockout pool preserves the heterogeneity of gene-editing outcomes without selection of a single clone, providing a versatile tool for functional studies.
The parental MCF-7 cell line is an epithelial, estrogen receptor-positive (ER+) breast adenocarcinoma model derived from the pleural effusion of a patient with metastatic breast cancer. Classified as the luminal A molecular subtype, MCF-7 cells retain functional estrogen receptor alpha (ER??) signaling and are widely employed in breast cancer research to model hormone-dependent tumor growth and metastasis. The well-characterized genetic landscape of MCF-7 cells offers a defined background in which to interrogate the contribution of JAG2 to oncogenic phenotypes.
JAG2 encodes a transmembrane Notch ligand that engages NOTCH1 and NOTCH3 receptors on adjacent cells. Receptor activation triggers sequential cleavage by ADAM10 and ??-secretase, releasing the Notch intracellular domain (NICD). NICD translocates to the nucleus and forms a complex with CSL (RBPJ) and MAML co-activators to drive transcription of targets including HES1, HEY1, MYC, CCND1, and BCL2. This signaling promotes proliferation and suppresses differentiation. JAG2 expression is regulated by TP53, HIF1A, NICD, and ER??, linking it to Wnt/??-catenin and PI3K/AKT/mTOR pathways.
In MCF-7 cells, JAG2-driven Notch signaling contributes to tumorigenic properties such as enhanced proliferation, survival, and invasive capacity. Disruption of JAG2 in this ER+ breast cancer model is predicted to impair NICD generation and attenuate expression of pro-proliferative and anti-apoptotic Notch targets. Consequently, this knockout model provides a valuable platform to dissect the role of JAG2 in hormone-responsive breast cancer, including its potential crosstalk with ER?? signaling and its contribution to the tumor?Cstroma interaction via Notch pathway modulation.
Applications include qRT-PCR for JAG2, HES1, and HEY1, Western blot analysis of Notch receptors and NICD, Notch luciferase reporter assays, and flow cytometry for surface JAG2. Phenotypic assays such as MTS/BrdU proliferation, Annexin V apoptosis, and Transwell migration/invasion can be performed, along with co-culture Notch activity assays. These tools support investigation of Notch signaling, breast cancer progression, drug target validation, and tumor?Cstroma interactions. For further information, contact Ascent Research.
