The ARFIP2 Knockout HAP1 Polyclonal Cells product consists of a polyclonal population of HAP1 cells edited by CRISPR/Cas9 to disrupt the ARFIP2 gene. This knockout model provides a loss-of-function system for studying ARFIP2-dependent processes without relying on transient suppression methods. The polyclonal format allows researchers to assess gene function in a mixed genetic background, which can help mitigate clone-specific artifacts. The disruption is designed to impair ARFIP2 protein function, enabling downstream analyses of its role in actin cytoskeleton dynamics and cell motility.
HAP1 cells are a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) line. They maintain the BCR-ABL1 translocation, rendering them sensitive to imatinib, and exhibit a fibroblast-like adherent morphology. The haploid nature reduces functional redundancy, making HAP1 a powerful platform for genetic screens and mechanistic studies.
ARFIP2 (also known as POR1) acts as a key effector downstream of the small GTPase ARF6. Upon activation, ARF6-GTP directly recruits ARFIP2, which in turn promotes RAC1 activation and binds cortactin through its SH3 domain. This complex stimulates localized actin polymerization, leading to membrane ruffling and enhanced cell migration. ARFIP2 thus bridges ARF6 signaling to actin remodeling, with upstream inputs from EGFR and PIP2, and downstream impacts on F-actin and cortactin.
In the HAP1 leukemia background, ARFIP2 knockout provides a unique tool to dissect ARF6-driven motility pathways that may contribute to the invasive phenotype of cancer cells. Because HAP1 cells retain BCR-ABL1 signaling, this model also permits investigation of potential crosstalk between oncogenic kinase pathways and ARF6-mediated cytoskeletal rearrangements, which is relevant to mechanisms of leukemia dissemination and solid tumor metastasis.
Typical research applications include wound healing migration assays, transwell invasion assays, and phalloidin staining to visualize F-actin changes. Researchers can also perform immunofluorescence for cortactin and membrane ruffles, ARF6 activity pull-downs, and co-immunoprecipitation of ARFIP2 complexes. This knockout model is suited for live-cell imaging of membrane protrusion dynamics and for screening regulators of metastasis. For more information, please contact Ascent Research.
