The ITGB3BP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population featuring disruption of the ITGB3BP gene. This loss-of-function model is derived from the HAP1 cell line and provides a versatile resource for investigating the roles of the calcium- and integrin-binding protein CIB1 in signal transduction, adhesion, and apoptosis. As a polyclonal pool, the cells encompass diverse gene-disrupted alleles, reducing clonal bias and enabling robust phenotypic analysis.
HAP1 is a near-haploid human hematopoietic cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) isolate. Its near-haploid karyotype minimizes genetic complexity, enhancing the efficiency of CRISPR-mediated knockout and simplifying genotype?Cphenotype correlations. The CML origin retains disease-relevant features, such as active integrin and survival signaling pathways, making it an ideal host for studying leukemogenesis and targeted therapy resistance.
ITGB3BP encodes the calcium-binding protein CIB1, which interacts directly with the cytoplasmic domain of integrin ??3 (ITGB3) to regulate integrin activation. CIB1 is activated by calcium (Ca2+) mobilization downstream of thrombin and vascular endothelial growth factor (VEGF) signaling. It subsequently modulates the activity of focal adhesion kinase (FAK), AKT, and ERK, while also controlling apoptotic effectors Caspase-3 and p21. CIB1 forms complexes with p21-activated kinase 1 (PAK1), cell division cycle 42 (CDC42), and GIPC1, connecting integrin and growth factor receptor signaling to the actin cytoskeleton and transcriptional regulation.
In the context of HAP1 cells, ITGB3BP knockout disrupts central integrin signaling nodes that are often dysregulated in CML. This model allows detailed examination of how CIB1-dependent adhesion and migration affect leukemic cell behavior, including proliferation, survival, and drug response. The near-haploid background facilitates clear dissection of the PI3K-Akt and FAK pathways, and the polyclonal nature ensures that observed phenotypes are not artifacts of a single clone. Consequently, this system is well-suited for studying the molecular basis of metastasis and thrombosis.
Key applications include functional genomics studies, integrin signaling pathway analysis, and apoptosis research. Representative assays include western blotting and RT-qPCR for gene expression profiling, cell adhesion and migration assays, Caspase-3-based apoptosis detection, and integrin activation flow cytometry. Co-immunoprecipitation can be performed to map CIB1 interaction networks. The polyclonal knockout pool is also valuable for drug target validation screens. For inquiries regarding this product or custom engineering services, please contact Ascent Research.
