The ARHGEF16 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9?edited polyclonal population of HAP1 cells carrying targeted disruption of the ARHGEF16 gene. This heterogeneous knockout pool enables robust loss?of?function studies without clonal selection artifacts, providing a reliable model to investigate ARHGEF16?dependent pathways in a near?haploid human background.
HAP1 is a near?haploid, adherent cell line derived from the male chronic myeloid leukemia (CML) line KBM?7. Its haploidy simplifies genetic analysis, as single?allele disruption yields complete loss of function. HAP1 cells are widely employed in functional genomics and high?throughput screening, and they retain key signaling pathways relevant to cancer cell motility and adhesion.
ARHGEF16 is a guanine nucleotide exchange factor (GEF) that activates RhoG by catalyzing GDP/GTP exchange. Active RhoG stimulates Rac1 and Cdc42, leading to PAK and LIMK kinase activation, cofilin phosphorylation, and actin polymerization required for lamellipodia formation and cell migration. This cascade is regulated by upstream signals from receptor tyrosine kinases, integrins, PI3K, Src kinases, and GPCRs, and it often involves cooperation with ELMO?DOCK scaffold complexes to control localized actin remodeling.
In HAP1 cells, ARHGEF16 knockout disrupts RhoG?Rac1/Cdc42 signaling, impairing cytoskeletal dynamics and attenuating migration and invasion. The haploid genome ensures unambiguous gene inactivation, making this model ideal for dissecting ARHGEF16??s role in motility. Given its CML origin, the model is particularly relevant for studying Rho GTPase signaling in hematologic cancers and metastasis, with implications for colorectal, breast, and glioblastoma malignancies.
These polyclonal knockout cells are suitable for a variety of functional assays, including scratch wound healing, transwell invasion, F?actin immunofluorescence, RhoG activation pull?downs, and western blotting for phospho?PAK and phospho?cofilin. Live?cell imaging and Rac1/Cdc42 activity assays can further quantify migration dynamics. The population format also supports high?throughput genetic or pharmacological screens targeting Rho?driven pathways. For technical information, please contact Ascent Research.
