ANXA3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the annexin A3 (ANXA3) gene. This knockout model is generated in the HAP1 cell line, providing a loss-of-function system to study ANXA3-dependent processes. The polyclonal format represents a heterogeneous pool of edited alleles, enabling robust phenotypic analysis without single-cell cloning. This product is suitable for investigating the role of ANXA3 in signal transduction, membrane dynamics, and disease-relevant pathways.
HAP1 is a near-haploid human fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype simplifies gene editing, as disruption of a single allele is sufficient to achieve complete knockout. HAP1 cells express the BCR-ABL oncogene and exhibit adherent growth, making them compatible with standard culture conditions and a wide range of functional assays. This genetic background is widely used for genetic screens and mechanistic studies due to its ease of manipulation and consistent phenotype.
ANXA3 encodes a calcium-dependent phospholipid-binding protein that regulates exocytosis, phagocytosis, cell migration, proliferation, and apoptosis. The protein is activated by upstream signals such as TNF??, IL-6, hypoxia (HIF-1??), and calcium ionophore. It interacts with phosphatidylserine, S100A10, integrin ??v??3, and protein kinase C to modulate membrane-associated signaling. Downstream, ANXA3 promotes AKT phosphorylation, ERK1/2 activation, and NF-??B p65 signaling, leading to VEGF expression and MMP2 secretion. Additionally, ANXA3 influences Bcl-2 family proteins, thereby controlling caspase-3-mediated apoptosis.
In HAP1 cells, ANXA3 knockout abolishes calcium-dependent phospholipid binding, disrupting membrane-proximal signal transduction and impairing NF-??B and PI3K/AKT pathways. This results in reduced phosphorylation of AKT and ERK1/2, diminished proliferation, impaired migration, and increased apoptosis. The haploid background ensures penetrant knockout phenotypes, as no wild-type allele compensates for the loss of ANXA3 function. Consequently, this model provides a clean genetic system for dissecting the cellular roles of ANXA3.
These polyclonal knockout cells are ideal for functional genomics, cancer cell biology, and drug target validation. Researchers can use western blotting and RT-qPCR to confirm ANXA3 depletion and assess downstream targets like VEGF and MMP2. Wound-healing assays measure migration, while Annexin V staining and MTS/MTT assays quantify apoptosis and proliferation. Immunofluorescence and phospho-AKT ELISA enable visualization of protein localization and signaling activity. For further information, contact Ascent Research.
