The ANXA5 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population available for functional studies of the ANXA5 gene in a human near-haploid background. This product delivers a heterogeneous pool of cells with targeted disruption of ANXA5, enabling loss-of-function analyses without the need for clonal isolation. The polyclonal format retains genetic diversity while providing a robust model for high-throughput screening and pooled knockout experiments.
HAP1 cells are a near-haploid human chronic myeloid leukemia cell line derived from the male KBM-7 blast crisis isolate. Adapted for both adherent and suspension culture, these cells maintain a predominantly haploid karyotype, which simplifies genetic manipulation by eliminating the need for homozygous targeting. This feature makes HAP1 a powerful platform for CRISPR-based functional genomics, as single-allele disruption readily generates complete knockout phenotypes, reducing experimental variability in pooled screens.
ANXA5 encodes a calcium-dependent phospholipid-binding protein that acts as a critical regulator of coagulation and apoptotic cell clearance. Mechanistically, ANXA5 binds phosphatidylserine on activated platelets and apoptotic cells in the presence of calcium ions, competitively inhibiting factor Xa and thereby dampening thrombin generation. It also bridges phosphatidylserine-exposing apoptotic cells to phagocytes via interaction with integrin ??v??5, promoting TGF-?? release and anti-inflammatory signaling. Upstream, ANXA5 expression is modulated by apoptotic stimuli such as TNF?? and staurosporine, glucocorticoids, and the REST transcription factor, while downstream it suppresses NF-??B and induces IL-10, reinforcing immune tolerance.
In the HAP1 context, loss of ANXA5 disrupts key pathways governing coagulation, efferocytosis, and inflammation. The near-haploid background ensures a straightforward genotype-phenotype correlation, facilitating the dissection of ANXA5??s role in diseases such as recurrent pregnancy loss, venous thromboembolism, pre-eclampsia, atherosclerosis, and various cancers. Moreover, this knockout model supports investigations into how ANXA5 deficiency alters cellular responses to external stimuli, providing a clean system to study pathway components like caspase 3/7, tissue factor, and TGF-?? without confounding diploid fluctuations.
Researchers can employ these cells in apoptosis recognition studies using flow cytometry for phosphatidylserine exposure, coagulation inhibition assays measuring factor Xa activity, and efferocytosis co-culture experiments quantifying TGF-?? release via ELISA. The polyclonal pool is well suited for CRISPR phenotypic screens, evaluating anti-inflammatory drug candidates, and cancer cell clearance models that probe interactions with MERTK and TIM-4. Western blotting and RT-qPCR further enable validation of downstream targets such as IL-10 and NF-??B. For further information and technical support, please contact Ascent Research.
