The ASPH Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the aspartate beta-hydroxylase (ASPH) gene in a human cervical cancer background. This product introduces CRISPR/Cas9-mediated gene disruption across the ASPH locus in a heterogeneous pool of HeLa cells, generating a versatile polyclonal knockout model that avoids clonal selection artifacts. The population is suitable for investigating ASPH-dependent signaling, proliferation, migration, and invasion phenotypes without the need for single-cell cloning, enabling robust and statistically representative functional analyses.
HeLa cells are an immortalized epithelial cell line derived from a cervical adenocarcinoma, widely employed as a model of human cervical cancer. These cells contain integrated human papillomavirus type 18 (HPV18) sequences, which contribute to their transformed phenotype and rapid growth. The HeLa background provides a physiologically relevant context for studying oncogenic signaling pathways, including Notch and HIF-1?? cascades, and is a standard host for gene-editing studies aimed at dissecting molecular mechanisms of tumorigenesis and metastatic progression.
ASPH encodes aspartate beta-hydroxylase, an enzyme that catalyzes the hydroxylation of aspartate and asparagine residues within EGF-like repeats of certain proteins, including Notch receptors (Notch1, Notch2) and their ligands (Jagged1, Jagged2). This post-translational modification enhances calcium binding and promotes Notch pathway activation, particularly under hypoxic conditions where ASPH is transcriptionally upregulated by HIF-1??. Activated Notch signaling drives the expression of downstream targets such as HES1, HEY1, c-Myc, and cyclin D1, thereby promoting cell proliferation, migration, and invasion. ASPH interacts with Fe2+, 2-oxoglutarate, and calcium ions, and its activity is modulated by upstream factors like insulin-like growth factor 1 (IGF-1) and miR-486.
In HeLa cells, ASPH-mediated hydroxylation of Notch receptors and Jagged ligands amplifies Notch signaling, contributing to the aggressive malignant phenotype. Disruption of ASPH in this knockout population attenuates Notch activation, reducing HES1 and c-Myc expression, and impairing cellular processes critical for cervical cancer progression, such as migration and invasion through extracellular matrix. This model thus allows direct interrogation of the ASPH?CNotch axis in a hypoxic tumor microenvironment, offering insights into the interplay between HIF-1?? stabilization, calcium homeostasis, and metastatic competency.
The ASPH Knockout HeLa Polyclonal Cells are suited for diverse research applications, including cancer biology, Notch signaling pathway analysis, hypoxia research, and drug discovery for ASPH inhibitors. Functional genomics studies can be performed using assays such as Western blotting, RT-qPCR, Notch luciferase reporter assays, Transwell migration/invasion assays, immunofluorescence for NICD, co-immunoprecipitation, HIF-1?? stabilization assays, calcium imaging, and RNA-seq. This knockout model empowers mechanistic investigations into ASPH-dependent metastasis mechanisms and serves as a platform for testing small-molecule inhibitors targeting the ASPH?CNotch interaction. For further technical details, please contact Ascent Research.
