The ASAP1 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population designed to disrupt the ASAP1 (ArfGAP with SH3 domain, ankyrin repeat, and PH domain) gene in the near-haploid HAP1 cell line. This product provides a pooled knockout model, avoiding single-cell cloning to maintain population diversity, enabling robust functional genomics studies and screening approaches that rely on averaged phenotypes across multiple editing events.
The HAP1 host cell line is a near-haploid (n = 24) human male fibroblast-like cell line derived from chronic myeloid leukemia KBM-7 cells. Its hemizygous genomic configuration simplifies CRISPR-mediated knockout generation and eliminates the complexity of functional redundancy, making it widely adopted for CRISPR-based functional genomics and knockout screening campaigns while preserving key adhesion and growth factor signaling machineries.
ASAP1 functions as a GTPase-activating protein (GAP) for Arf family small GTPases, primarily Arf1 and Arf6, catalyzing GTP hydrolysis to inactivate these molecular switches. It is recruited to integrin adhesion complexes and activated by upstream signals from epidermal growth factor (EGF), platelet-derived growth factor (PDGF), and SRC family kinases. ASAP1 physically interacts with focal adhesion kinase (FAK), PYK2, cortactin, paxillin, and Crk-associated substrate (CAS). Its GAP activity toward Arf6 promotes cortical actin remodeling through cortactin, linking integrin and receptor tyrosine kinase signaling to focal adhesion turnover, actin polymerization, and cell migration.
In HAP1 cells, the hemizygous nullizygous disruption of ASAP1 eradicates its GAP function, leading to sustained Arf6 activation and consequent dysregulation of actin cytoskeleton dynamics and focal adhesion organization. This near-haploid model provides a clean genetic background to dissect ASAP1-dependent pathways driving cell motility, invasion, and metastatic behavior, with direct relevance to studies on breast cancer progression, melanoma invasion, and malignant transformation.
Researchers can validate target disruption with Sanger sequencing, confirm loss of protein expression via western blotting and immunofluorescence, and assess transcript levels by RT-qPCR. Functional assays include wound healing migration assays, Transwell invasion assays, cell adhesion assays, and immunofluorescence imaging of actin stress fibers and focal adhesion markers. Biochemical interaction assays such as co-immunoprecipitation with ASAP1 partners (e.g., SRC, FAK, cortactin) and direct Arf-GAP activity measurements further enable mechanistic studies. These polyclonal knockout cells are optimized for research into cancer metastasis, cytoskeletal dynamics, and Arf signaling. For technical inquiries, please contact Ascent Research.
