The CCR2 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population targeting the CCR2 gene in the human near-haploid HAP1 cell line. This pooled population ensures disruption of CCR2 function across cells, establishing a loss-of-function model for investigating CCR2-dependent pathways. The polyclonal format maintains genetic diversity while minimizing clonal artifacts, making it suitable for functional genomics and pathway analyses.
HAP1 cells are a near-haploid chronic myeloid leukemia-derived line with a fibroblast-like morphology and a haploid karyotype, which streamlines gene targeting as single-allele disruption yields a functional knockout. Originating from KBM-7 cells, HAP1 is extensively used in genetic perturbation screens and functional genomics due to its stable karyotype and amenability to CRISPR/Cas9 editing and lentiviral transduction, enabling efficient generation of knockout populations.
CCR2 is a G-protein coupled receptor for the chemokine CCL2 (MCP-1) and other ??-chemokines, including CCL7, CCL8, CCL12, and CCL13. Ligand binding activates G??i proteins, stimulating phospholipase C (PLC), calcium mobilization, and MAPK/ERK and PI3K-Akt signaling cascades, which drive chemotaxis, integrin activation, and pro-inflammatory cytokine expression (e.g., IL-6, TNF-??). Upstream regulators such as TNF-??, IL-1??, and NF-??B enhance CCR2 expression, while downstream effectors include MAPK/ERK, JNK, PI3K, Akt, and NF-??B. The receptor interacts with ??-arrestin, GRK2/3, and HIV co-receptors CD4 and CCR5, placing it at the intersection of inflammatory signaling, immune cell trafficking, and viral entry mechanisms.
In the HAP1 haploid background, CCR2 knockout abolishes CCL2-induced chemotactic and signaling responses, enabling clear dissection of CCR2-dependent pathways without diploid genetic complexity. This model is ideal for studying mechanisms of monocyte migration, inflammation, and signal transduction, and for conducting genetic screens to identify modifiers of chemokine receptor activity. The polyclonal nature reduces clonal bias, providing a robust platform for comparative functional studies.
Typical applications include transwell migration and microfluidic chemotaxis assays, calcium flux measurements, phospho-ERK/Akt western blotting, RT-qPCR for cytokine gene expression, flow cytometry-based integrin activation assays, co-immunoprecipitation of receptor complexes, and cAMP inhibition assays. These cells support research into inflammatory diseases (atherosclerosis, rheumatoid arthritis, multiple sclerosis), HIV co-receptor biology, cancer metastasis, and monocyte/macrophage function, and are suitable for drug target validation and functional genomics screening. For further information or to discuss custom applications, please contact Ascent Research.
