ARHGAP12 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HAP1 human near-haploid chronic myeloid leukemia cell line. This product is designed for loss-of-function studies of the ARHGAP12 gene, which encodes a Rho GTPase-activating protein. The polyclonal knockout pool provides a heterogeneous population of edited cells, enabling robust investigation of ARHGAP12-dependent cellular processes without the need for single-cell clonal isolation.
The HAP1 cell line is derived from a chronic myeloid leukemia patient and features a predominantly haploid karyotype, which simplifies genetic manipulation and facilitates high-throughput genetic screens. These cells exhibit a fibroblast-like morphology and are widely used for studying haploid cell biology and performing CRISPR-based functional genomics. The near-haploid background reduces genetic redundancy, enhancing the penetrance of gene disruption phenotypes.
ARHGAP12 functions as a GTPase-activating protein (GAP) for the Rho family GTPases Rac1 and Cdc42, accelerating GTP hydrolysis and thereby inactivating these molecular switches. It acts downstream of growth factors, integrin signaling, and receptor tyrosine kinases, and interacts with p120-catenin (CTNND1), Src, and focal adhesion kinase (FAK). ARHGAP12 negatively regulates actin polymerization, cell migration, and adhesion dynamics through modulation of downstream effectors such as PAK, LIMK, cofilin, and the Arp2/3 complex. Its activity impacts cytoskeletal organization, focal adhesion turnover, and cellular protrusion formation.
In the HAP1 cell context, disruption of ARHGAP12 provides a powerful model to dissect Rho GTPase signaling pathways involved in leukemic cell migration, adhesion, and cytoskeletal reorganization. Because HAP1 cells are inherently haploid, a single allelic disruption is sufficient to achieve functional knockout, making this polyclonal population particularly suitable for phenotypic screens and quantitative assays. This model is relevant for investigating mechanisms underlying cancer cell invasion, neurological disorders, and cardiovascular diseases where Rho GTPase dysregulation is implicated.
Typical applications include Western blotting for Rho GTPase activity using phospho-specific or pull-down assays, immunofluorescence staining of actin filaments and focal adhesion markers (e.g., vinculin, paxillin), time-lapse microscopy for migration and invasion, co-immunoprecipitation to probe protein interactions, and G-LISA or other GTPase activation assays. Additionally, these cells can be employed in arrayed or pooled CRISPR phenotypic screens to identify genetic interactions. For additional technical information, please contact Ascent Research.
