The CCL7 Knockout HAP1 Polyclonal Cells provide a ready-to-use CRISPR/Cas9-edited polyclonal knockout cell population in which the CCL7 gene has been disrupted. This loss-of-function model eliminates CCL7 protein expression, facilitating the study of chemokine-mediated cellular responses. The polyclonal pool retains biological variation and is well-suited for high-throughput screening and genetic interaction studies.
The parental HAP1 cell line, derived from the KBM-7 chronic myeloid leukemia line, is a human near-haploid cell model widely used in functional genomics. HAP1 cells grow as an adherent monolayer and exhibit a stable near-haploid karyotype, simplifying knockout generation and ensuring that gene disruptions are effectively homozygous. This genetic simplicity makes HAP1 an ideal background for CRISPR-based genetic screens and pathway dissection.
CCL7 (monocyte chemoattractant protein-3) is a potent chemokine that orchestrates leukocyte migration. It signals through the G-protein-coupled receptors CCR1, CCR2, and CCR3, leading to calcium mobilization and activation of downstream effectors including MAPK1, MAPK3, AKT, and STAT3. Its expression is transcriptionally regulated by pro-inflammatory stimuli such as TNF-alpha, IL-1, and IFN-gamma via NF-??B and AP-1. CCL7 also binds glycosaminoglycans and is processed by MMP2, which modulates chemokine availability and gradient formation, ultimately directing the chemotaxis of monocytes, eosinophils, basophils, and T lymphocytes.
In the HAP1 near-haploid background, knockout of CCL7 eliminates its chemotactic activity, offering a clean loss-of-function system to investigate inflammatory cell recruitment. This model is highly relevant for diseases such as asthma, atherosclerosis, rheumatoid arthritis, and cancer metastasis, where CCL7-driven leukocyte trafficking contributes to pathogenesis. The absence of a second gene copy in HAP1 cells ensures complete penetrance of the knockout phenotype, enhancing the reliability of drug target validation and signaling studies.
Common applications include quantitative RT-qPCR and Western blotting to confirm knockout, Boyden chamber chemotaxis assays to measure monocyte migration, and ELISA-based profiling of secreted chemokines. Phospho-signaling analysis of MAPK1/3 and AKT further dissects intracellular pathways. The cells can be employed in high-throughput chemical screens, RNA-seq, flow cytometry, and monocyte adhesion assays. This polyclonal knockout cell population is a versatile tool for inflammation, immunology, and oncology research. For additional information, contact Ascent Research.
