The CCL7 Knockout HCT 116 Polyclonal Cells are a heterogeneous CRISPR/Cas9-edited population derived from the HCT 116 human colorectal carcinoma cell line, designed to disrupt the CCL7 gene and eliminate expression of the monocyte chemoattractant protein-3 (MCP-3). This polyclonal knockout model provides a loss-of-function tool for studying CCL7-mediated processes without the biases introduced by clonal selection, ensuring a broad representation of genetic backgrounds. The cells are supplied as a ready-to-use pool suitable for immediate expansion and downstream functional assays, enabling robust and reproducible investigation of chemokine biology.
The HCT 116 host cell line is an adherent epithelial line established from a human colorectal carcinoma, characterized by microsatellite instability (MSI-H) and a well-documented activating KRAS mutation. These genetic features make HCT 116 a widely used model for cancer signaling, tumor microenvironment studies, and drug screening. The cell line??s epithelial origin and colorectal cancer context are particularly relevant for exploring chemokine-mediated communication between tumor cells and immune components, as well as for assessing how oncogenic pathways intersect with inflammatory signaling networks.
CCL7, a member of the CC chemokine family, functions as a potent chemoattractant for monocytes, eosinophils, basophils, and T cells by binding to the G protein-coupled receptors CCR1, CCR2, CCR3, and CCR5. This interaction activates downstream cascades involving calcium flux, PI3K/AKT, MAPK/ERK, and NF-??B, which collectively drive cell migration, survival, and inflammatory gene expression. CCL7 expression is transcriptionally upregulated by pro-inflammatory stimuli including TNF, IL-1??, IFN-??, and TLR4 ligands such as LPS, linking its production to innate immune responses. Once secreted, CCL7 interacts with glycosaminoglycans to form immobilized gradients on endothelial surfaces, facilitating directed leukocyte trafficking. Disruption of CCL7 in this model thus interrupts a critical node in chemokine signaling and immune cell recruitment.
In the context of HCT 116 colorectal cancer cells, ablation of CCL7 offers a powerful system to dissect autocrine and paracrine chemokine circuits that may influence tumor progression, immune evasion, and metastatic potential. Because HCT 116 cells harbor an oncogenic KRAS mutation, the knockout model enables investigation of cross-talk between Ras-driven pathways and chemokine expression, a key area in inflammation-associated carcinogenesis. Researchers can explore how loss of CCL7 alters the secretion of other cytokines, modifies activation of NF-??B and MAPK/ERK, and impacts the recruitment of immune cells in co-culture or conditioned medium transfer experiments, providing insights into the tumor microenvironment??s immunosuppressive or pro-inflammatory characteristics.
Typical applications include chemotaxis assays to quantify immune cell migration toward conditioned media, ELISA and multiplex analysis of secreted factors, and western blot or flow cytometry to assess signaling pathway activation (e.g., phospho-ERK, phospho-AKT). The polyclonal knockout cells are well-suited for tumor immunology research, drug target validation against inflammatory or metastatic processes, and functional studies of CCL7-dependent gene regulation using RT-qPCR. They can also be employed in co-culture systems to model the interaction between tumor-derived chemokines and immune cells, aiding in the development of therapeutic strategies that modulate the chemokine milieu. For additional information, please contact Ascent Research.
