The ASPH Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, featuring targeted disruption of the ASPH gene. This loss-of-function model is designed for investigating the biological roles of aspartate ??-hydroxylase (ASPH) in a genetically tractable near-haploid system.
HAP1 cells are a near-haploid, adherent, fibroblast-like cell line originally derived from the KBM-7 chronic myeloid leukemia line. The near-haploid karyotype facilitates functional genomics and genetic screening applications, as recessive phenotypes can be uncovered in a single-step gene disruption. HAP1 cells maintain stable growth and are amenable to many routine cell-based assays, making them a robust platform for studying signaling pathways and cellular processes.
ASPH catalyzes hydroxylation of aspartic acid and asparagine residues in EGF-like domains of Notch receptors and ligands, thereby potentiating Notch pathway activation. Its expression is positively regulated by HIF1A and negatively by ING4 and the miR-200 family. ASPH directly engages NOTCH1, JAG1, and DLL4 in a calcium-dependent manner. Downstream, it enhances the expression of Notch target genes HES1 and HEY1 and activates SRC and FAK to drive cell migration and invasion. Core pathway components including ADAM10 and PSEN1 mediate the proteolytic activation of Notch receptors, highlighting the central role of ASPH in modulating both receptor-ligand interactions and downstream signaling outputs.
In the HAP1 near-haploid background, ASPH knockout phenotypes are not masked by a second functional allele, providing a clear window into ASPH-dependent functions. This model enables precise dissection of ASPH??s impact on Notch receptor maturation, ligand activation, and the downstream transcriptional program, as well as on cell migration and invasion capacities, in a genetically simplified context.
This knockout cell product is ideally suited for functional characterization of Notch signaling, cancer invasion and metastasis research, and drug target validation. Researchers can employ western blotting to confirm ASPH ablation, RT-qPCR to quantify Notch target gene expression, and migration or invasion assays to assess phenotypic changes. Co-immunoprecipitation enables analysis of ASPH interactions with NOTCH1 or its ligands, while phospho-signaling analysis evaluates SRC/FAK activation. Immunofluorescence and Notch reporter assays provide additional spatial and transcriptional readouts. For further details, please contact Ascent Research.
