BIN3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the BIN3 gene has been disrupted by non-homologous end joining-mediated repair. This product contains a heterogeneous mixture of edited HAP1 cells, providing a robust loss-of-function model for population-level assays. The polyclonal format avoids clonal selection biases and captures the diversity of CRISPR-induced mutations, making it ideal for assays requiring average gene disruption effects across a cell population.
The HAP1 host is a near-haploid human cell line derived from the chronic myeloid leukemia KBM-7 line, which harbors the BCR-ABL1 fusion oncogene characteristic of CML. Its haploid karyotype, with a single copy of most chromosomes, simplifies gene targeting and reduces genetic redundancy, facilitating straightforward knockout interpretation. HAP1 cells are male, adherent, and widely adopted in high-throughput screening and cancer research, offering a well-characterized platform for studying gene function in a leukemic context.
BIN3 encodes a BAR domain-containing protein that functions as a membrane curvature sensor and scaffold, essential for clathrin-mediated endocytosis and actin cytoskeleton remodeling. BIN3 is activated downstream of epidermal growth factor receptor (EGFR) and integrin-mediated adhesion and interacts directly with Dynamin-2, Cortactin, Clathrin, and the AP-2 complex. It couples membrane invagination to actin polymerization by integrating signals from the EGFR-Ras-Rac1/Cdc42 axis to activate the Arp2/3 complex. Through these interactions, BIN3 facilitates both the formation and scission of endocytic vesicles, regulating the internalization of receptor tyrosine kinases and other cargo.
In the HAP1 CML background, BIN3 knockout provides a powerful system for investigating how BAR domain proteins influence oncogenic signaling. Disruption of BIN3 is expected to impair clathrin-mediated endocytosis, potentially altering the trafficking and signaling of surface receptors such as EGFR and integrins. This may modulate downstream pathways including Ras-MAPK and actin remodeling, relevant to leukemia cell proliferation and migration. The near-haploid genomic context further enables clean genotype-phenotype correlations, making this model valuable for synthetic lethality screens and drug target identification.
Representative applications include endocytosis assays using transferrin or EGF uptake, membrane trafficking studies via confocal immunofluorescence, and drug target validation through Western blot and flow cytometry. Co-immunoprecipitation experiments can probe BIN3-containing complexes, while cell migration assays assess cytoskeletal phenotypes. Researchers can quantify receptor internalization kinetics, evaluate signal transduction changes via phospho-specific antibodies, and screen for genetic interactions using CRISPR libraries. For further technical details, please contact Ascent Research.
