The JAG2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-mediated polyclonal knockout population engineered to disrupt the JAG2 gene in the HAP1 human near-haploid cell line. This loss-of-function model enables targeted study of Jagged-2-dependent Notch signaling without the constraints of monoclonal selection, maintaining cellular heterogeneity relevant to population-level analyses. The polyclonal format provides a versatile tool for investigating ligand-driven Notch activation in a genetically tractable background.
HAP1 cells are derived from the KBM-7 chronic myeloid leukemia (CML) line and stably maintain a near-haploid karyotype, which greatly facilitates gene editing and functional genomics applications. Originally established for haploid genetic screens, these cells retain leukemic characteristics and active signaling pathways, making them a well-characterized system for knockout studies in cancer biology and hematopoiesis research.
JAG2 encodes Jagged-2, a DSL family transmembrane ligand that activates Notch receptors (NOTCH1?C4) on adjacent cells. Upon receptor?Cligand engagement, sequential proteolysis by ADAM10 and the ??-secretase complex releases the Notch intracellular domain (NICD), which translocates to the nucleus and forms a transcriptional complex with RBPJ/CSL and Mastermind-like (MAML) coactivators. This complex drives expression of canonical targets such as HES1, HES5, HEY1, HEY2, MYC, and CCND1, thereby orchestrating cell fate decisions, proliferation, and apoptosis. Upstream regulators including NF-??B, E-protein transcription factors (TCF3/E2A), and TGF-?? modulate JAG2 expression, while MIB1 ubiquitin ligase fine-tunes ligand activity. Thus, JAG2 serves as a critical node in juxtacrine signaling, linking extracellular cues to nuclear Notch outputs.
Disruption of JAG2 in the HAP1 background is particularly informative for dissecting Notch ligand function in the context of CML and hematopoietic disorders. Because HAP1 cells harbor active Notch signaling components and are amenable to high?throughput screening, the knockout population allows researchers to distinguish between canonical ligand?dependent signaling and non?canonical or ligand?independent Notch activation. This system supports the study of developmental pathways deregulated in leukemia and can reveal synthetic lethal interactions or resistance mechanisms relevant to Notch?targeted therapies.
The JAG2 Knockout HAP1 Polyclonal Cells are suited for a broad array of experimental applications, including functional characterization of Notch signaling in cancer, haploid genetic screens to identify modulators of the pathway, and validation of potential drug targets. Researchers can employ techniques such as western blotting for Notch pathway proteins, RT?qPCR for HES1, flow cytometry to assess surface Notch expression, co?immunoprecipitation of JAG2?CNotch interactions, and Notch reporter luciferase assays. Additional utility includes apoptosis assays, migration/invasion studies, and drug sensitivity profiling. For technical support or further information, please contact Ascent Research.
