The APMAP Knockout HAP1 Polyclonal Cells provide a heterogeneous CRISPR/Cas9-edited population of HAP1 cells harboring targeted disruption of the APMAP gene. This pooled knockout model is an ideal tool for loss-of-function investigations of APMAP in a near-haploid human cell background. By eliminating functional APMAP protein expression, the cells enable dissection of APMAP-dependent processes in integrin biology, adipocyte signaling, and cancer cell adhesion.
These cells are generated in the HAP1 host cell line, a near-haploid human cell model originally derived from the KBM-7 chronic myeloid leukemia (CML) line. The haploid genome facilitates efficient CRISPR/Cas9 editing and reduces genetic redundancy, making HAP1 a robust platform for knockout studies. Retaining key features of leukemic cells, HAP1 provides a relevant context for exploring signaling pathways that drive proliferation and survival in CML and other cancers.
APMAP encodes a transmembrane O-glycosyltransferase that specifically modifies integrin ??1 (ITGB1), a critical regulator of cell?Cextracellular matrix adhesion. Under the control of pro-adipogenic transcription factors PPAR?? and C/EBP??, APMAP glycosylates ITGB1, modulating its affinity for ligands and downstream signaling via focal adhesion kinase (FAK) and SRC. APMAP-dependent glycosylation promotes phosphorylation of FAK and subsequent activation of ERK1/2 and AKT pathways, linking extracellular cues to cytoskeletal reorganization and transcriptional responses. The enzyme functions in complex with integrin ??V and ITGB1, and its loss abrogates FAK-mediated signal transduction, thereby impairing cell adhesion, migration, and adipogenic commitment.
In HAP1 cells, which endogenously express integrins and exhibit adhesion-dependent growth, APMAP disruption offers a clean system to dissect ITGB1 glycosylation and its functional consequences. The knockout cells are expected to display reduced adhesion, altered migration, and attenuated FAK/SRC/ERK/AKT signaling, recapitulating phenotypes observed in APMAP-deficient models. Because HAP1 is a leukemic line, this model is particularly valuable for studying how integrin glycosylation influences cancer cell dissemination and for screening therapeutic interventions targeting adhesion pathways.
Typical applications include quantitative cell adhesion and migration assays, western blotting for phospho-FAK and phospho-ERK, lectin blotting to assess integrin glycosylation status, and flow cytometry to monitor surface ITGB1 expression. The knockout cells also support adipogenic differentiation studies when supplemented with appropriate stimuli, with lipid accumulation visualized by Oil Red O staining. This product is suitable for both mechanistic studies and high-throughput screens. For inquiries and technical support, please contact Ascent Research.
