The CCL2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the CCL2 gene has been disrupted within the human HAP1 cell line. This polyclonal pool provides a loss-of-function model for studying CCL2-dependent signaling and function in a near-haploid genetic background, avoiding clonal artifacts and enabling robust phenotypic screening.
The HAP1 cell line is a human haploid fibroblast-like line derived from the KBM-7 chronic myeloid leukemia lineage. Its near-haploid karyotype simplifies genetic knockout generation, as disruption of a single allele is sufficient to ablate gene function without diploid compensation. HAP1 cells exhibit adherent, fibroblastoid morphology and are widely used in CRISPR-based functional genomics, high-content imaging, and pooled perturbation screens. Their stable growth and compatibility with lentiviral transduction make them an ideal platform for chemokine pathway interrogation.
CCL2 (MCP-1) is a chemokine that directs monocyte, memory T cell, and dendritic cell migration by activating the cognate receptor CCR2. Receptor engagement triggers G??i-dependent activation of PI3K/AKT, MAPK/ERK, and JAK/STAT cascades. CCL2 expression is transcriptionally regulated by TNF-??, IL-1??, IFN-??, and LPS through NF-??B and AP-1. Downstream effectors include MMP-9, IL-6, and MCPIP1, as well as integrin activation and reactive oxygen species generation. Extracellular modulators such as ACKR2 and glycosaminoglycans fine-tune chemokine gradients, while heparin binding influences ligand presentation. Thus, CCL2 sits at a nexus of inflammatory cytokine networks and leukocyte transendothelial migration.
In this knockout model, CRISPR/Cas9-mediated disruption of CCL2 abrogates autocrine/paracrine CCL2-CCR2 signaling, resulting in loss of PI3K/AKT, MAPK/ERK, and JAK/STAT pathway activation. This eliminates CCL2 secretion and downstream inflammatory gene expression, providing a clean background for dissecting CCL2-dependent phenotypes. The haploid context ensures that gene disruption on a single allele yields a functional null state in the bulk population. Consequently, the cells are adept at revealing roles of CCL2 in monocyte chemotaxis, migration, and invasion, and are especially valuable for paracrine signaling studies in co-culture settings with immune cells.
The CCL2 Knockout HAP1 Polyclonal Cells support a wide range of experimental applications, including chemotaxis assays, ELISA measurement of CCL2, and western blotting for phospho-AKT and phospho-ERK. They facilitate pooled CRISPR screens for CCL2-independent migration factors and drug target validation with CCR2 antagonists or PI3K inhibitors. In cancer immunology, co-culture with monocytes allows dissection of tumor-immune crosstalk, while NF-??B p65 translocation assays interrogate inflammatory dynamics. RT-qPCR profiling and live-cell imaging of leukocyte recruitment are also readily performed. For additional details, please contact Ascent Research.
