The CCL23 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the human CCL23 gene has been disrupted. This product offers a genetically heterogeneous pool of cells carrying loss-of-function mutations across the CCL23 locus, providing a robust model system for interrogating the biological functions of this chemokine without the limitations of single-clone variability. The polyclonal format enables researchers to study CCL23-dependent phenotypes in a near-haploid background, facilitating large-scale functional genomics and signaling analyses. As with all polyclonal knockout products, individual cells may harbor distinct editing outcomes, collectively yielding a comprehensive loss-of-function effect suitable for pooled assays and replicate experiments.
The HAP1 host cell line is a near-haploid human cell model derived from the KBM-7 chronic myeloid leukemia line. HAP1 cells exhibit adherent growth with fibroblast-like morphology and retain a single copy of most chromosomes, except for a disomic region on chromosome 8. This near-haploid genomic configuration simplifies genetic analyses, as a single allelic disruption is sufficient to abolish gene function. The male origin and stable karyotype make HAP1 a widely adopted system for CRISPR-based loss-of-function screens, drug-sensitivity profiling, and mechanistic pathway studies.
CCL23, a CC chemokine also known as myeloid progenitor inhibitory factor 1, engages the CCR1 receptor to trigger G??i-linked signaling cascades. Receptor activation leads to PI3K-dependent phosphorylation of Akt and ERK1/2, which in turn promote RhoA GTPase activity, integrin-mediated adhesion, cell migration, and the release of pro-inflammatory cytokines including IL-6 and IL-8. Expression of CCL23 is upregulated by TNF-alpha, IL-1beta, and LPS via NF-kappaB, while its availability is fine-tuned by the decoy receptor ACKR2 and glycosaminoglycan binding. RGS proteins accelerate G?protein inactivation, adding another layer of regulation. Collectively, this signaling network integrates immune cell chemotaxis with inflammatory and metastatic processes.
The HAP1 near-haploid background ensures that phenotypes observed in CCL23 knockout cells are directly attributable to gene disruption, enabling unambiguous dissection of chemokine-driven signaling. Researchers can assess CCL23-dependent effects on ERK/Akt phosphorylation, cell migration, and cytokine secretion, and validate upstream activators like TNF-alpha or downstream mediators such as RhoA. This clean genetic platform is ideally suited for modeling pathologies linked to CCL23, including rheumatoid arthritis, multiple sclerosis, and metastasis.
Typical applications of these polyclonal knockout cells include chemotaxis and invasion assays, phospho-ERK/Akt analysis by flow cytometry or Western blotting, and co-immunoprecipitation of CCR1 signaling complexes. They are also suited for RNA-seq transcriptomics and high-content screening in inflammation and cancer research, making them a versatile tool for functional genomics, drug target validation, and immunology studies. For additional technical specifications or ordering inquiries, please contact Ascent Research.
