The DISP3 Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited, polyclonal knockout cell population for loss-of-function analysis of DISP3. This heterogeneous pool of HAP1 cells features CRISPR/Cas9-mediated disruptions at the DISP3 locus, abolishing functional protein expression without single-cell cloning. The polyclonal format is ideal for rapid functional genomic screens and pooled assays, minimizing clonal artifacts.
The HAP1 cell line is a near-haploid, adherent human cell model derived from a male chronic myeloid leukemia (CML) patient. Despite its hematopoietic origin, HAP1 exhibits a fibroblast-like morphology and retains signaling competency. Its near-haploid karyotype ensures efficient CRISPR/Cas9 gene disruption, as targeting a single allele yields a complete loss-of-function phenotype, making it a preferred system for genetic screens and mechanistic studies.
DISP3 encodes a dispatched protein that mediates the release of cholesterol-modified Hedgehog ligands??SHH, IHH, and DHH??from producing cells. This secretion process involves interactions with cholesterol and the SCUBE2 cofactor. Released ligands engage the PTCH1 receptor on receiving cells, derepressing SMO and activating GLI transcription factors (GLI1, GLI2, GLI3). Negative regulators include HHIP and PTCH1. DISP3 therefore governs the extracellular availability of Hedgehog morphogens, critically shaping signaling gradients.
In the HAP1 background, DISP3 knockout permits rigorous dissection of Hedgehog pathway dynamics and ligand secretion. The model is invaluable for studying developmental disorders and cancers linked to Hedgehog misregulation, such as basal cell carcinoma and medulloblastoma. By eliminating DISP3 function, researchers can probe how loss of ligand release influences downstream signaling and assess tumor cell dependency on Hedgehog paracrine communication.
This polyclonal knockout pool facilitates diverse experimental workflows, including CRISPR screens, qRT-PCR and Western blot analyses of GLI1 and PTCH1, luciferase-based pathway activity assays, and immunofluorescence for ligand distribution. Cell migration assays and drug sensitivity testing with Hedgehog inhibitors (e.g., vismodegib) are also enabled. The polyclonal population is particularly suited for large-scale functional studies. For further information, please contact Ascent Research.
