The DISP1 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 human cell line. This product is designed to disrupt the Dispatched 1 (DISP1) gene, generating a loss-of-function model for studying Hedgehog signaling and associated cellular processes. The polyclonal format provides a heterogeneous pool of edited cells, enabling robust and scalable genetic analysis without requiring single-cell cloning.
HAP1 is a human near-haploid fibroblastoid cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) line. It carries the Philadelphia chromosome and exhibits a predominantly haploid karyotype, making it exceptionally suited for functional genomics and knockout studies due to the unambiguous genotype-phenotype correlations. The near-haploid nature ensures that a single genetic alteration is directly reflected at the phenotypic level, which is particularly advantageous for dissecting signal transduction pathways.
DISP1 encodes a transmembrane transporter essential for the secretion of cholesterol-modified Hedgehog ligands, including Sonic Hedgehog (SHH), Indian Hedgehog (IHH), and Desert Hedgehog (DHH). In the canonical Hedgehog pathway, DISP1 functions downstream of ligand production and upstream of receptor binding; it interacts with SCUBE2 and heparan sulfate proteoglycans to facilitate the release of dually lipidated Hedgehog proteins. Upon release, these ligands engage the receptor Patched 1 (PTCH1), relieving suppression of Smoothened (SMO), which activates GLI transcription factors (GLI1, GLI2) that regulate expression of targets such as CCND1, BCL2, HHIP, and FOXA2. Disruption of DISP1 abolishes paracrine Hedgehog signaling, as the producing cells fail to secrete active ligand, thus preventing pathway activation in receiving cells.
In the HAP1 background, the DISP1 knockout provides a clean genetic tool to interrogate Hedgehog pathway dependency without confounding effects from paralogous genes or redundant signaling routes, thanks to the haploid genome. This model is particularly relevant for research into developmental disorders linked to disrupted Hedgehog signaling, such as holoprosencephaly, midline craniofacial defects, and congenital heart disease. By eliminating DISP1 function in a near-haploid system, researchers can directly link genotype to pathway output, facilitating drug target validation and modifier screening.
This polyclonal knockout pool is ideally suited for a range of functional assays, including GLI luciferase reporter assays to measure pathway activity, RT-qPCR for GLI1 and PTCH1 transcript levels, and western blotting to quantify GLI1 protein. Conditioned media from edited cells can be assessed by SHH ELISA to confirm impaired ligand secretion, while immunofluorescence staining for primary cilia can reveal structural or trafficking defects. Cell viability assays further enable compound sensitivity profiling. Additionally, the product supports genetic interaction screens and high-throughput target validation studies. For additional information, please contact Ascent Research.
