The CCL20 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population featuring targeted disruption of the CCL20 gene in the near-haploid HAP1 cell line. This knockout model enables loss-of-function studies of CCL20, a chemokine critical for immune cell recruitment, without the complexities of clonal isolation. The polyclonal nature provides a heterogeneous knockout population suitable for pool-based genetic screens and bulk functional assays.
HAP1 cells are a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, featuring a fibroblast-like adherent morphology and BCR-ABL expression. The haploid karyotype simplifies genetic analysis and reduces functional redundancy, making HAP1 an ideal background for CRISPR-based knockout screening. The lack of a second allele in most chromosomes ensures that a single CRISPR-induced mutation can generate a complete loss-of-function phenotype in the majority of cells, facilitating straightforward genotype-phenotype correlations.
CCL20 is a chemoattractant cytokine that orchestrates immune cell trafficking by binding to the CCR6 receptor, a G??i-coupled GPCR. Expression of CCL20 is strongly induced by proinflammatory stimuli, including TNF-??, IL-1??, and IL-17, through activation of transcription factors such as NF-??B and C/EBP??. Upon CCL20 engagement, CCR6 triggers downstream signaling via G??i, PLC, PI3K, Akt, and ERK1/2, leading to calcium mobilization, cytoskeletal rearrangement, and directed migration of CCR6-expressing cells such as Th17 lymphocytes and immature dendritic cells. The CCL20-CCR6 axis thus plays a pivotal role in immune surveillance, inflammatory responses, and the tumor microenvironment.
This knockout model is particularly valuable for studying CCL20-CCR6 signaling in inflammation and cancer. By ablating CCL20, researchers can assess its role in immune cell recruitment and validate the CCL20-CCR6 axis as a therapeutic target. The HAP1 host’s near-haploid genome ensures that gene disruption is complete in most cells, enabling robust detection of phenotypic changes in pooled populations. This polyclonal knockout approach facilitates high-throughput screening for inhibitors or modulators of the CCL20-CCR6 pathway, as well as systematic analysis of CCL20 regulatory networks and downstream effector mechanisms.
Typical applications include high-throughput screening of chemokine inhibitors using ELISA-based detection of secreted CCL20, mechanistic dissection of NF-??B-driven CCL20 transcription via RT-qPCR and Western blotting, and chemotaxis assays to measure functional consequences of CCL20 loss on CCR6-expressing cell migration. The polyclonal knockout cells also enable pooled RNA-seq studies to identify downstream transcriptional targets and pathway perturbations, supporting systems-level analysis of the CCL20-CCR6 signaling network. For further technical details or to place an order, please contact Ascent Research.
