The DLG1 Knockout HAP1 Polyclonal Cells are a validated CRISPR/Cas9-edited polyclonal knockout cell population targeting the DLG1 gene in HAP1 haploid human cells. This heterogeneous loss-of-function model contains diverse gene disruptions and is ideal for pooled functional screens and population-based phenotypic assays, compatible with both arrayed and pooled screening formats. The polyclonal format minimizes clonal selection artifacts, ensuring robust biological representation.
HAP1 is a near-haploid human cell line derived from a chronic myelogenous leukemia patient, widely used as a model for genetic perturbation studies. Its haploid karyotype enables efficient CRISPR/Cas9-mediated gene disruption with only one allele targeted. HAP1 cells retain many epithelial signaling pathways and are frequently employed in functional genomics, drug target discovery, and cancer biology research, offering fast growth and compatibility with diverse in vitro assays.
DLG1 encodes a multi-PDZ domain scaffold protein that organizes adherens junctions and controls cell polarity. It physically links E-cadherin to the actin cytoskeleton, serving as a nexus for Hippo and Wnt signaling. Upstream regulators include E-cadherin, Frizzled receptors, AMPK, Aurora kinase A, and Nedd4 ligases. DLG1 scaffolds the ??-catenin destruction complex and Hippo kinase module, interacting with APC, PTEN, and MST1/2?CLATS1/2 to control YAP/TAZ, and modulates AKT and ERK to affect cell cycle progression. Complexes with LIN7, CASK, and DLGAP1 further integrate tight junction and synaptic plasticity pathways.
In the HAP1 haploid background, DLG1 knockout disrupts cell?Ccell adhesion and apical?Cbasal polarity, leading to enhanced migration and proliferation. This model recapitulates features of colorectal and gastric cancers and is relevant to inflammatory bowel disease, where DLG1 loss compromises epithelial barrier integrity. The haploid karyotype simplifies genotype?Cphenotype correlations, allowing unambiguous attribution of functional defects to DLG1 disruption. HAP1 cells retain endogenous Hippo and Wnt signaling machinery, providing an ideal context for mechanistic studies. Additionally, the polyclonal nature of the knockout population enables robust detection of subtle phenotypes in pooled assays and screens.
Applications include cell adhesion and migration assays (wound healing), Wnt/??-catenin reporter studies (TOP/FOP flash luciferase), epithelial barrier function analyses, and drug target validation. Compatible techniques cover western blotting, co-immunoprecipitation, immunofluorescence microscopy, RNA-seq, and flow cytometry for cell cycle analysis. The polyclonal knockout population is well-suited for pooled functional screens and robust pharmacological profiling. For further technical details or custom inquiries, please contact Ascent Research.
