The CCR10 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population originating from the HAP1 human haploid cell line, engineered for targeted disruption of the CCR10 gene. This loss-of-function model abolishes CCR10-mediated signaling, providing a defined system to investigate chemokine receptor function in a near-haploid genetic background. The polyclonal composition ensures a heterogeneous pool of edited cells, circumventing clonal selection biases and offering a robust platform for functional assays.
HAP1 is a chronic myeloid leukemia-derived human cell line with a near-haploid karyotype, derived from the KBM-7 clone. Its haploid nature facilitates unambiguous genotype-phenotype correlations in genetic studies, particularly for recessive mutations. HAP1 cells exhibit rapid doubling times, stable growth, and a well-characterized signaling network, making them an optimal host for CRISPR/Cas9-mediated gene disruption experiments targeting chemokine receptors and downstream effector pathways.
CCR10 encodes a G-protein-coupled receptor selectively activated by the chemokine ligands CCL27 and CCL28. Ligand engagement triggers G??i-mediated inhibition of adenylate cyclase, reducing intracellular cAMP, and promotes calcium mobilization. This initiates phosphorylation cascades involving ERK and AKT through the MAPK/ERK and PI3K-Akt pathways, respectively. CCR10 signaling also facilitates integrin activation, driving chemotaxis and adhesion of T cells. Regulatory factors such as beta-arrestins and GNAI proteins modulate receptor desensitization and signal termination. Thus, CCR10 functions as a central node in skin-homing T cell trafficking, coupling chemotactic cues to cytoskeletal and transcriptional responses.
In HAP1 cells, CCR10 disruption eliminates ligand-induced signaling outputs, creating a clean background for pathway dissection. The haploid genotype ensures that a single functional knockout is sufficient to observe phenotypes, simplifying data interpretation. This polyclonal knockout population mirrors the genetic diversity found in typical editing experiments, allowing researchers to assess pooled cellular responses without clonal artifacts. It is particularly suited for high-throughput screening of compounds targeting CCR10 or its downstream effectors, as well as for studying receptor crosstalk in a minimized signaling environment.
This knockout model is ideal for chemotaxis and transwell migration assays to assess CCR10-dependent migration, calcium flux measurements for G-protein-coupling analysis, and western blot detection of phospho-ERK and phospho-AKT. Flow cytometry and RT-qPCR confirm knockout fidelity and downstream transcriptional changes, while adhesion assays examine integrin activation. Applications span inflammatory skin diseases such as psoriasis and atopic dermatitis, cutaneous T-cell lymphoma, and drug target validation. The polyclonal knockout cells provide a robust resource for exploring skin-homing T cell biology. Contact Ascent Research for additional details.
