The CCL24 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered for targeted disruption of the human CCL24 gene. This product provides a heterogeneous pool of knockout cells, avoiding clonal heterogeneity and enabling robust functional studies without the constraints of single-cell-derived lines. The polyclonal format ensures representation of the full knockout spectrum, making it ideal for assays that require population-level responses, such as chemotaxis or signaling analyses. The gene disruption is achieved through CRISPR/Cas9-mediated non-homologous end joining, which introduces loss-of-function mutations across the cell population. Researchers can utilize these cells as a ready-to-use knockout model for dissecting CCL24-dependent biology in a controlled genetic background.
The host cell line, HAP1, is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) model with a male karyotype. Its near-haploid genome simplifies genetic manipulation and interpretation, as only one allele is typically present, reducing the likelihood of heterozygous editing outcomes. HAP1 cells are widely adopted for haploid genetic screens and functional genomics due to their stable growth characteristics and absence of diploid masking effects. The CML origin provides a hematopoietic background relevant to leukocyte signaling and chemokine biology, making it a suitable chassis for studying CCL24 function in the context of immune cell behavior and inflammatory responses.
CCL24, also known as eotaxin-2, encodes a CC chemokine that signals exclusively through the CCR3 receptor, a G-protein-coupled receptor expressed on eosinophils, basophils, and Th2 lymphocytes. Ligand binding triggers G??i-mediated signaling, leading to activation of effectors such as phospholipase C (PLC), which generates IP3 and promotes calcium flux, and the MAPK/ERK and PI3K/AKT cascades. These pathways drive cytoskeletal rearrangement via actin polymerization and direct cell migration. Upstream, CCL24 expression is transcriptionally regulated by IL-4, IL-13, and STAT6, while TNF-?? and IL-5 further potentiate its production. CCL24 also interacts with glycosaminoglycans for tissue localization and is scavenged by the atypical chemokine receptor ACKR2 (D6). In this knockout model, disruption of CCL24 eliminates autocrine or paracrine signaling through this axis, providing a null background for functional reconstitution studies.
In the HAP1 context, CCL24 knockout cells serve as a powerful tool for dissecting CCR3-mediated signaling networks without interference from endogenous ligand. The near-haploid genetics of HAP1 ensure that the targeted disruption is effectively hemizygous, minimizing residual gene expression. Loss of CCL24 abolishes CCR3-dependent calcium mobilization, ERK phosphorylation, and cell migration, as outlined in the mechanistic summary. This clean knockout system is particularly valuable for studying eosinophil biology and allergic inflammation, where CCL24 is a key chemoattractant. The cells can be used to validate the specificity of anti-CCR3 therapeutics or to overexpress mutant CCL24 variants for structure?Cfunction analyses. Furthermore, the model allows exploration of compensatory chemokine networks in the absence of eotaxin-2.
Typical research applications include functional genomics screening with arrayed libraries, where the CCL24 knockout population serves as a foundational line for synthetic lethality or genetic interaction mapping. In drug discovery, these cells are suitable for high-throughput chemotaxis assays to screen CCR3 antagonists, with readouts such as transwell migration or impedance-based methods. Calcium flux assays using fluorescent dyes (e.g., Fluo-4) and flow cytometric detection of activation markers are also representative protocols. Electrophoretic and gene-expression analyses (Western blot, RT-qPCR) confirm knockout status and downstream effector activation, while ELISA quantifies secreted chemokines. The polyclonal nature supports robust statistical analysis across replicates. For further information or technical support, please contact Ascent Research.
