CCL5 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the CCL5 gene in the near-haploid HAP1 cell line. The polyclonal format comprises a heterogeneous pool of edited cells, each carrying CRISPR/Cas9-mediated gene disruption of CCL5, and provides a practical loss-of-function model without requiring single-cell clone isolation. This product is designed for researchers needing to interrogate CCL5-dependent processes in a uniform genetic background, offering a robust tool for chemokine signaling studies, drug screening, and functional genomics.
The HAP1 host cell line is a near-haploid human cell model originally derived from the KBM-7 chronic myeloid leukemia (CML) line. Its near-haploid karyotype simplifies genetic manipulation, as most genes exist in a single copy, enabling efficient knockout generation and clear phenotypic analysis. HAP1 cells maintain core signaling pathways relevant to hematopoietic biology and cancer, making them a versatile platform for investigating cytokine networks, immune cell migration, and intracellular signal transduction.
CCL5 (C-C motif chemokine ligand 5, also called RANTES) is a potent chemoattractant cytokine that orchestrates the recruitment of T cells, monocytes, and eosinophils to sites of inflammation. Its expression is induced by upstream regulators including TNF-alpha, IL-1beta, and IFN-gamma, acting through transcription factors such as NF-kappaB and IRF3. Secreted CCL5 engages chemokine receptors CCR1, CCR3, and CCR5, as well as the co-receptor DPP4 and glycosaminoglycans, to initiate G-protein-mediated signaling. Downstream cascades include PI3K-Akt, JAK2-STAT3, and MAPK/ERK1/2 pathways, which drive calcium mobilization and activate Rho GTPases. Transcriptional outcomes involve upregulation of matrix metalloproteinases (MMPs), adhesion molecules ICAM-1 and VCAM-1, and pro-inflammatory cytokines IL-6 and IL-8, thereby promoting immune cell adhesion, migration, and tissue invasion.
In the HAP1 background, CCL5 knockout eliminates these signaling inputs, creating a clean genetic system to dissect chemokine-driven mechanisms. The near-haploid nature of the host cells ensures that loss-of-function phenotypes are unambiguous, facilitating direct interpretation in genetic screens and targeted biochemical assays. This model is particularly valuable for studying CCR5-mediated events, given that CCL5 is a primary endogenous ligand for CCR5??a co-receptor for HIV entry and a validated target in inflammatory diseases and cancer metastasis. Researchers can use this knockout pool to examine how the absence of CCL5 alters downstream kinase activity, receptor expression patterns, and cross-communication with parallel chemokine pathways.
Typical research applications include functional characterization of CCL5 in chemokine signaling, chemotaxis and migration assays, compound screening for CCR5 antagonists, HIV co-receptor studies, and investigations into inflammation and tumor cell migration. Compatible experimental techniques encompass Western blotting for signaling intermediates such as phospho-ERK1/2, phospho-Akt, and phospho-STAT3; RT-qPCR for cytokine expression profiling; transwell migration assays; flow cytometry for surface receptor quantification; ELISA for secreted CCL5; phospho-kinase activity measurements; reporter assays for NF-kappaB or STAT3; RNA-seq for transcriptome-wide analysis; and co-immunoprecipitation for receptor-ligand interactions. For further information or technical support, please contact Ascent Research.
