The ITGB1BP1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ITGB1BP1 gene in HAP1 cells, providing a loss-of-function model for studying integrin ??1 binding protein 1 (ICAP-1). This polyclonal population allows pooled analysis of gene disruption effects without clonal isolation, suitable for investigating ITGB1BP1-dependent mechanisms in cell adhesion, migration, and vascular signaling.
HAP1 cells are a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia background, characterized by a single copy of most chromosomes except a diploid portion of chromosome 15. This ploidy simplifies CRISPR/Cas9-based knockout generation, as target gene disruption in a single allele is sufficient to achieve functional knockout, making HAP1 a widely used host for genetic studies. The cells retain key signaling machinery relevant to adhesion and cytoskeletal regulation, providing a consistent genetic background for interrogating gene function.
ITGB1BP1 encodes ICAP-1, which binds the cytoplasmic tail of integrin ??1 and acts as a negative regulator of integrin activation, thereby inhibiting cell adhesion and migration. ICAP-1 is activated downstream of integrin ??1 engagement and focal adhesion kinase (FAK) signaling, and it directly interacts with KRIT1 (CCM1), recruiting it to the plasma membrane. This interaction bridges integrin- and cerebral cavernous malformation (CCM) complex signaling, with ICAP-1 modulating the CCM complex comprising KRIT1, CCM2, and PDCD10 (CCM3). Mechanistically, ITGB1BP1 restricts RhoA-ROCK signaling, thereby controlling actin stress fiber formation, focal adhesion dynamics, and actomyosin contractility. Additional interactions with RAP1A further link ITGB1BP1 to integrin-mediated cytoskeletal reorganization and endothelial barrier function.
In HAP1 cells, disruption of ITGB1BP1 is expected to derepress integrin ??1 activation and enhance cell-substrate adhesion, while altering RhoA-ROCK-dependent cytoskeletal tension. The haploid genetic background facilitates a clear loss-of-function phenotype, making these knockout cells a straightforward system to dissect ITGB1BP1’s role in adhesion-dependent signaling cascades. This model is particularly valuable for studying how integrin-CCM cross-talk regulates cellular morphology and motility, with potential implications for vascular integrity and cerebral cavernous malformation pathogenesis.
The ITGB1BP1 Knockout HAP1 Polyclonal Cells enable functional assays such as cell adhesion and spreading, wound healing migration, RhoA activation G-LISA, and co-immunoprecipitation to analyze interactions between ICAP-1, integrin ??1, and CCM complex components. Immunofluorescence reveals actin cytoskeleton and focal adhesion organization, while western blotting for phospho-FAK and flow cytometry for integrin activation states provide quantitative signaling readouts. These cells facilitate research into integrin-mediated adhesion, Rho GTPase signaling, cerebral cavernous malformation, cancer cell invasion, and vascular biology. Please contact Ascent Research for further details.
