The BRK1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human BRK1 gene. Generated from the HAP1 near-haploid cell line via CRISPR/Cas9-mediated gene disruption, this polyclonal pool provides a robust loss-of-function model while avoiding single-cell cloning artifacts. The product enables functional studies of BRK1-dependent actin dynamics and cell migration without the need for clonal isolation.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, exhibiting adherent fibroblast-like morphology and male origin. Its near-haploid karyotype simplifies genetic analysis and phenotypic interpretation, as most genes are present in a single copy. Widely used as a hematopoietic progenitor model and for leukemogenesis studies, HAP1 provides a relevant cellular context for exploring signaling networks governing cytoskeletal organization and adhesion.
BRK1 is an essential subunit of the heteropentameric WAVE regulatory complex (WRC), which also contains WASF1, CYFIP1, NCKAP1, and ABI1/2. The WRC transduces signals from Rac1 GTPase to the Arp2/3 complex, driving actin nucleation and polymerization at the leading edge of migrating cells. Rac1 activation by growth factors or integrins releases WRC inhibition of Arp2/3, promoting lamellipodial protrusion. BRK1 knockout disrupts WRC assembly, abrogating Rac1-induced actin polymerization and impairing interactions with actin monomers and ARPC1B.
In the HAP1 background, BRK1 disruption enables direct assessment of migration defects arising from WRC dysfunction within a leukemic cell context. The cell line??s endogenous expression of Rac1 and associated factors permits phenotypic linkage to upstream regulators such as NCKAP1 and ABI1/2. The near-haploid genome reduces genetic redundancy, improving genotype-phenotype correlations. This model is particularly suited to metastasis research, where BRK1-dependent lamellipodia contribute to invasion, and for studying actin cytoskeleton contributions to immunological disorders.
This polyclonal knockout product supports live-cell imaging of actin dynamics, wound healing and transwell migration assays, and immunofluorescence for F-actin and lamellipodia. Co-immunoprecipitation can verify WAVE complex integrity, and Rac1 pull-down assays delineate upstream signaling. The model is also amenable to drug response profiling and genetic interaction screens targeting the actin cytoskeleton. For further details or customization inquiries, contact Ascent Research.
