The CCL28 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-engineered heterogeneous population of HAP1 cells carrying targeted disruption of the CCL28 gene. As a polyclonal knockout product, this pool contains diverse gene-edited variants, enabling robust loss-of-function analyses without single-cell-clone constraints. The gene encodes the mucosae-associated chemokine CCL28, a key mediator of immune cell trafficking to mucosal surfaces. This product facilitates investigation of CCL28’s role in chemokine signaling and mucosal immunity.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) blast crisis isolate. The male-derived line retains a predominantly haploid karyotype, simplifying CRISPR/Cas9 editing and enhancing knockout generation efficiency. Although originating from a hematopoietic malignancy, HAP1??s haploid genetic makeup renders it a versatile platform for genetic screening, receptor pharmacology, and signal transduction studies. Its rapid proliferation and stable growth support high-throughput applications, including chemokine receptor functional assays and genome-wide knockout screens.
CCL28 functions as a chemokine that selectively binds the G-protein-coupled receptors CCR10 and, with lower affinity, CCR3. Upon ligand-receptor engagement, CCL28 activates G??i-dependent cascades, leading to downstream stimulation of PI3K and MAPK pathways. These events direct chemotaxis of IgA-secreting plasmablasts and T lymphocytes to mucosal epithelial barriers. CCL28 expression is upregulated by IL-4, IL-13, TLR ligands, and proinflammatory cytokines, integrating innate and adaptive immunity signals. Through CCR10-mediated chemotaxis, CCL28 orchestrates mucosal humoral responses and antimicrobial defense.
In HAP1 cells, CCL28 knockout provides a unique tool to dissect the CCL28?CCCR10/CCR3 signaling node independent of the endogenous chemokine milieu of primary immune cells. The haploid background minimizes genetic redundancy, facilitating unambiguous functional readouts. This model enables systematic assessment of receptor coupling, G-protein activation kinetics, and downstream PI3K/MAPK dynamics following exogenous chemokine stimulation. Additionally, the leukemic origin offers an opportunity to explore cross-talk between chemokine signals and oncogenic pathways, broadening relevance to tumor microenvironment research.
Researchers employ this polyclonal knockout population in chemotaxis and transwell migration assays to quantify CCL28-dependent cell recruitment, complemented by flow cytometry for CCR10 surface expression. ELISA and Western blotting validate chemokine level changes and signaling effector phosphorylation. The cells support antimicrobial activity testing and serve as a platform for inflammatory bowel disease modeling, where CCL28-mediated lymphocyte homing is pathologically relevant. For detailed characterization data and validation reports, please contact Ascent Research.
