The ARFIP1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population featuring disruption of the human ARFIP1 gene in the HAP1 cell line. This product provides a mixed population of edited cells with loss-of-function mutations in ARFIP1, enabling the study of membrane trafficking and actin remodeling without the constraints of single-cell clonal variation. The polyclonal format preserves genetic heterogeneity while ensuring robust knockout across the population, making it suitable for pooled functional assays and high-throughput screening applications.
HAP1 cells are a near-haploid human cell line derived from the chronic myeloid leukemia cell line KBM-7. With a haploid karyotype (except for a small portion of chromosome 15), HAP1 cells are widely used in functional genomics and knockout screens due to the ease of generating complete gene disruptions. As a hematopoietic progenitor cell model, they retain signaling pathways relevant to blood cell biology, yet their near-haploidy makes them an efficient platform for CRISPR/Cas9-based genetic engineering and subsequent phenotypic analyses.
ARFIP1 (ARF-interacting protein 1) is a critical effector of ADP-ribosylation factor (ARF) GTPases, particularly ARF1, ARF5, and ARF6. Through its BAR domain, ARFIP1 senses membrane curvature and orchestrates membrane tubulation, linking activated ARF proteins to actin cytoskeleton remodeling. ARFIP1 functions downstream of ARF activation by guanine nucleotide exchange factors, and its activity is modulated by phosphatidylinositol 4,5-bisphosphate (PIP2). ARFIP1 interacts directly with ARF GTPases and PIP2, and it regulates downstream targets including the actin cytoskeleton, Golgi membranes, endocytic vesicles, and Rac1 signaling. This positions ARFIP1 at the intersection of Arf6 signaling, membrane trafficking, and actin dynamics, with roles in Golgi structure maintenance and vesicle formation.
In HAP1 cells, disruption of ARFIP1 provides a unique model to dissect the interplay between membrane remodeling and the actin cytoskeleton in a near-haploid background. Because HAP1 cells are of hematopoietic origin, they express machinery relevant to cellular adhesion, migration, and endocytosis, processes often dysregulated in cancer metastasis. The knockout of ARFIP1 in these cells allows researchers to examine defects in Golgi organization, vesicle transport, and actin-based processes with reduced genetic redundancy. This model is particularly valuable for studying how ARF-dependent pathways control cell morphology and invasive behavior, and for identifying synthetic lethal interactions with other trafficking or cytoskeletal regulators.
Typical applications include high-content imaging to assess Golgi fragmentation and actin cytoskeleton reorganization via immunofluorescence, biochemical analysis of ARF-ARFIP1 interaction by co-immunoprecipitation, and functional assays such as transwell migration or actin polymerization assays. The polyclonal population is suitable for drug sensitivity screens to identify compounds that differentially affect wild-type versus ARFIP1-null cells, as well as for pooled CRISPR screens to study genetic interactions. Researchers can also utilize these cells for western blotting confirmation of protein loss and for investigating ARF-dependent signaling in a clean genetic background. For further product details, please contact Ascent Research.
