CCL3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the CCL3 gene in the near-haploid HAP1 human cell line. This loss-of-function model facilitates investigation of the chemokine CCL3 (MIP-1??) and its signaling roles. As a polyclonal preparation, it enables rapid functional assessment without single-cell cloning, making it suitable for high-throughput and preliminary screens. The cells are applicable to a range of biomedical research, including inflammation, oncology, and infectious disease.
HAP1 is a suspension-adapted human cell line with a predominantly haploid karyotype, originally derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid genome simplifies genetic manipulation, as disruption of a single allele often yields functional knockout. The cell line retains hematopoietic progenitor features and myeloid markers. Coupled with robust growth characteristics, HAP1 is widely used for CRISPR screening, signaling studies, and chemokine biology.
CCL3 (MIP-1??) is a CC chemokine chemoattractant for monocytes, T cells, and eosinophils, signaling through CCR1 and CCR5, G-protein-coupled receptors that activate G-alpha-i. Receptor engagement induces intracellular calcium flux and stimulates PI3K/Akt and MAPK/ERK phosphorylation cascades, leading to chemotaxis, cytoskeletal reorganization, and MMP-9 secretion. CCL3 expression is induced by TNF-??, IL-1??, and LPS via NF-??B and TLR4 pathways. CCL3 also binds glycosaminoglycans to form chemotactic gradients and acts as a CD4 ligand to enhance HIV-1 entry through CCR5. Key downstream effectors include focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (Pyk2), which regulate migration and adhesion.
In the HAP1 near-haploid background, CCL3 disruption produces a polyclonal pool with attenuated CCL3 function, allowing unambiguous signaling analysis without wild-type allele interference. This model is ideal for chemokine receptor pharmacology studies, as HAP1 cells express endogenous signaling components. The cells permit systematic dissection of PI3K/Akt and ERK pathway roles in migration, calcium responses, and inflammatory gene expression. With relevance to rheumatoid arthritis, HIV infection, multiple myeloma, and inflammatory bowel disease, this system supports disease mechanism studies and therapeutic screening.
Key applications include chemotaxis and calcium flux assays, flow cytometric analysis of CCR1/CCR5 expression, western blotting for phospho-ERK, and ELISA or RT-qPCR for CCL3 quantification. Co-immunoprecipitation can assess CCL3-CCR5 interactions, and the cells support drug target validation in immuno-oncology and anti-inflammatory research. Combining a tractable genetic background with a defined loss-of-function model, these cells offer a robust system for studying chemokine-driven signaling. For further technical details or to discuss custom applications, please contact Ascent Research.
