The JAG1 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout population derived from the human near-haploid HAP1 cell line, designed for loss-of-function studies of the JAG1 gene. This heterogeneous pool harbors diverse gene disruptions, providing a robust model for investigating JAG1-dependent pathways without clonal bias.
HAP1 is a near-haploid chronic myeloid leukemia cell line, originally derived from the KBM-7 line, with an adherent fibroblast-like morphology and a male origin. As a hematopoietic progenitor model, HAP1 retains features relevant to myeloid biology and offers a simplified genetic background that facilitates high-efficiency CRISPR-based knockout generation. The haploid state enables direct genotype-phenotype correlations, making it a preferred system for genetic screens and functional genomics.
JAG1 encodes a DSL domain-containing ligand that activates Notch receptors (NOTCH1-4) through direct cell?Ccell contact. Ligand-receptor engagement triggers sequential proteolysis by ADAM10/17 metalloproteases and gamma-secretase, releasing the Notch intracellular domain (NICD). NICD subsequently translocates to the nucleus and associates with RBPJ and MAML1 to drive transcription of key targets such as HES1, HEY1, MYC, cyclin D1, and CDKN1A. JAG1 activity is modulated by Fringe proteins (LFNG, MFNG, RFNG) and is essential for cell fate decisions, hematopoiesis, and angiogenesis.
Disruption of JAG1 in HAP1 cells provides a physiologically relevant platform to study Notch signaling in a human hematopoietic context. The near-haploid genome minimizes background genetic variability, enabling unambiguous assessment of JAG1??s role in progenitor cell proliferation, differentiation, and apoptosis. The polyclonal nature of the knockout population allows for the evaluation of a spectrum of allelic variations, simulating the heterogeneity observed in diseases such as Alagille syndrome and myeloid malignancies.
These cells are ideally suited for a variety of research applications, including Notch signaling pathway analysis via western blotting, RT-qPCR, RNA-seq, or luciferase reporter assays. They facilitate hematopoiesis and cancer studies through cell differentiation, proliferation, and apoptosis assays, and enable drug screening against Notch pathway components. Co-immunoprecipitation and flow cytometry can be employed to examine JAG1-interacting proteins and downstream effector expression. Additionally, the polyclonal format supports functional genomic screens and genetic interaction mapping. For further information, please contact Ascent Research.
