The ARHGAP39 Knockout HAP1 Polyclonal Cells comprise a population of HAP1 cells subjected to CRISPR/Cas9-mediated disruption of the ARHGAP39 gene, generating a heterogeneous pool of loss-of-function alleles within the host genome. This polyclonal knockout product provides a robust and scalable model for interrogating ARHGAP39-dependent signaling networks, bypassing the need for single-cell clone isolation. The resulting gene-edited cell population is suitable for pooled functional genomics, biochemical, and cell-based assays that require a near-complete ablation of target gene expression across the majority of cells.
The HAP1 cell line is a near-haploid, fibroblast-like derivative of the KBM-7 chronic myeloid leukemia cell line. This genetic background simplifies loss-of-function studies by eliminating a second functional allele, enabling rapid gene function assignment. HAP1 cells are widely used in genetic screens, drug target identification, and mechanistic cell biology, and their adherent, spread morphology suits studies of cell adhesion, migration, and cytoskeletal dynamics.
ARHGAP39 encodes a Rho GTPase-activating protein (GAP) that negatively regulates RhoA, Rac1, and Cdc42. It acts downstream of EphA4 receptor, activated by Src kinases and growth factor signals, and interacts with Nck and actin. By promoting GTP hydrolysis, ARHGAP39 attenuates ROCK and MLC phosphorylation, reorganizing the actin cytoskeleton, modulating focal adhesion turnover, and reducing cell migration and adhesion. This axis is integral to neurite outgrowth, cancer invasion, and morphogenesis.
Disruption of ARHGAP39 in HAP1 cells creates a platform to dissect EphA4?CRho GTPase cytoskeletal signaling. The haploid background ensures phenotypes are directly attributable to ARHGAP39 loss, avoiding compensatory effects. This model is advantageous for studying cancer cell migration and neurological disorders involving Rho GTPase signaling in axonal guidance and synaptic plasticity. The fibroblast-like morphology of HAP1 cells recapitulates mesenchymal migration and adhesion, making this knockout population a relevant tool for both basic and translational research.
These polyclonal cells support diverse applications: validation of CRISPR screens, biochemical analysis of EphA4?CARHGAP39?CRhoA modules via co-immunoprecipitation and immunofluorescence, RhoA activation assays, wound healing migration, phospho-MLC western blotting, and RT-qPCR profiling. High-content imaging of F-actin and focal adhesions is also feasible. For further details, contact Ascent Research.
