The AP2M1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the AP2M1 gene in the HAP1 human cell line. This loss-of-function model provides a heterogeneous gene-edited pool suitable for pooled population-based assays without the need for single-cell cloning, enabling robust functional studies.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) cell line. Its haploid genome simplifies genetic knockout analysis and offers a clean background for dissecting clathrin-mediated endocytosis, receptor signaling, and cancer cell biology. The leukemic origin also makes it relevant for investigating endocytic adaptor proteins in hematological malignancies.
AP2M1 encodes the ?? subunit of the adaptor protein complex 2 (AP-2), which recognizes tyrosine-based sorting signals (YXX??) on cargo receptors including transferrin receptor (TFRC), low-density lipoprotein receptor (LDLR), epidermal growth factor receptor (EGFR), and Notch. The ?? subunit interacts with clathrin heavy chain, AP2A1, AP2B1, epsin, Eps15, and HIP1R to assemble the endocytic coat at phosphatidylinositol-4,5-bisphosphate (PIP2)-rich plasma membrane domains. AP2M1 activity is regulated by receptor tyrosine kinases such as EGFR and by phosphorylated adaptors Dab2 and ARH. After vesicle budding, dynamin facilitates scission, followed by endosomal maturation through Rab5 and EEA1. Disruption of AP2M1 uncouples cargo recognition from coat assembly, impairing internalization of multiple receptors and altering downstream signaling and nutrient uptake.
In HAP1 leukemic cells, AP2M1 knockout disrupts clathrin-mediated endocytosis of EGFR and LDLR, attenuating oncogenic signaling and cholesterol homeostasis??processes linked to familial hypercholesterolemia and cancer. The model further enables study of viral entry pathways that depend on clathrin, and its haploid status allows unambiguous genotype-phenotype associations.
This polyclonal knockout population supports a broad range of applications including high-content screening for endocytosis modulators, transferrin uptake assays, EGFR degradation kinetics by western blotting and immunofluorescence, flow cytometric quantification of surface receptor levels (e.g., TFRC, LDLR), and co-immunoprecipitation of AP-2 complex components. Researchers may also perform cell proliferation assays and phospho-signaling analysis to evaluate downstream effects. For additional technical information, please contact Ascent Research.
