The ARPC5L Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human ARPC5L gene in the HAP1 cell line. This mixed-pool product provides a heterogeneous collection of loss-of-function alleles, enabling functional studies without clonal selection effects. It facilitates investigation of ARPC5L roles in actin cytoskeleton remodeling, cell motility, and endocytosis. As a polyclonal knockout, it offers population-level gene disruption suitable for assays where average gene ablation is required. Researchers can use this system to dissect the contribution of ARPC5L to Arp2/3 complex activity in a haploid genetic background.
The HAP1 host cell line is a near-haploid human myeloid leukemia cell line derived from KBM-7 cells, originally isolated from a chronic myeloid leukemia patient. The haploid karyotype simplifies CRISPR/Cas9-mediated gene disruption, as targeting a single allele yields a loss-of-function phenotype, eliminating the need for biallelic inactivation. HAP1 cells are widely used in functional genomics and high-throughput genetic screens due to this feature. They grow adherently and maintain key cancer-relevant signaling pathways, including those regulating actin dynamics, making them an appropriate model for ARPC5L knockout studies.
ARPC5L encodes a subunit of the Arp2/3 complex, the primary nucleator of branched actin networks. The complex is activated by nucleation-promoting factors such as N-WASP and the WAVE complex, themselves stimulated by Rho GTPases Rac1 and Cdc42, and by PIP2. ARPC5L interacts with all core Arp2/3 subunits (ARP2, ARP3, ARPC1-4) and associates with cortactin and WIP. This machinery drives actin polymerization, generating lamellipodia and filopodia-like protrusions, and powers endocytic vesicle trafficking and cell adhesion. Consequently, ARPC5L is critical for cell migration and intracellular transport.
In HAP1 cells, loss of ARPC5L disrupts Arp2/3 complex integrity, leading to aberrant actin organization and impaired cell motility, endocytosis, and adhesion. Given the leukemic origin of HAP1, this model is particularly relevant for examining how actin remodeling contributes to cancer cell invasion and metastasis. The adherent culture format permits high-resolution fluorescence imaging of F-actin using phalloidin, allowing direct visualization of cytoskeletal defects. The polyclonal knockout population assesses the overall functional impact without the adaptation biases of single-cell clones.
These polyclonal knockout cells are suitable for a range of assays. Western blotting and RT-qPCR can verify ARPC5L depletion, while immunofluorescence microscopy reveals altered F-actin organization. Migration and invasion can be measured via transwell or scratch assays. Co-immunoprecipitation experiments can probe Arp2/3 complex assembly. Applications span cancer metastasis research, where the cells help clarify the role of branched actin in tumor dissemination, and drug discovery, screening for modulators of actin dynamics. For additional information and ordering, please contact Ascent Research.
