HSPG2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population carrying targeted disruption of the HSPG2 gene in the HAP1 near-haploid human cell line. This knockout model abolishes expression of the large heparan sulfate proteoglycan perlecan, a central basement membrane component, enabling systematic dissection of extracellular matrix (ECM) biology and growth factor signaling in a simplified genetic background. The polyclonal nature of the knockout pool ensures representation of diverse editing events within the target locus, providing a versatile loss-of-function system for functional genomics and pathway analysis.
The HAP1 cell line is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, retaining a single copy of most chromosomes except for a disomy of chromosome 8. These fibroblast-like, adherent cells exhibit a stable near-haploid karyotype that facilitates CRISPR-based gene disruption and phenotype?Cgenotype correlation studies. The reduced genetic redundancy of HAP1 cells amplifies the impact of monogenic perturbations, making them ideally suited for high-content screening and robust functional validation of target genes such as HSPG2.
Perlecan, encoded by HSPG2, is a large heparan sulfate proteoglycan that anchors basement membranes by binding collagen IV, laminin, fibronectin, and nidogen, and it engages cell surface integrins and dystroglycan. It functions as a co-receptor for heparin-binding growth factors including FGF2, VEGF, and PDGF, facilitating their presentation to tyrosine kinase receptors and subsequent activation of downstream FAK, ERK1/2, and AKT signaling. HSPG2 expression is induced by TGFB1, TNF, and IL1B via NF-??B and SMAD pathways. Knockout of HSPG2 abrogates perlecan scaffolding, decoupling growth factor bioavailability from integrin-mediated mechanotransduction and attenuating ERK and AKT phosphorylation.
In the near-haploid HAP1 background, the HSPG2 knockout eliminates perlecan-mediated ECM organization and growth factor presentation without the masking effects of a diploid ECM proteoglycan redundancy. This genetic clarity enhances the detection of subtle phenotypes in cell adhesion, focal adhesion dynamics, and signal transduction, and renders the model particularly advantageous for arrayed or pooled CRISPR modifier screens. By combining the HSPG2 disruption with the streamlined HAP1 genome, researchers can interrogate perlecan??s role in controlling integrin?CFAK?CERK and PI3K?CAKT pathways with minimal confounding variables, establishing a clean baseline for rescue experiments and drug response profiling.
These polyclonal knockout cells are designed for western blot analysis of perlecan and phospho-ERK/AKT, immunofluorescence of focal adhesions, cell adhesion and migration assays, and growth factor binding studies. They enable investigation of ECM deposition and perlecan interactome via co-immunoprecipitation, as well as disease modeling for Schwartz-Jampel syndrome and cancer, and drug screening targeting ECM?Cintegrin?Ckinase pathways. For ordering or technical inquiries, contact Ascent Research.
