CCR5 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal population of the near-haploid human HAP1 cell line in which the CCR5 gene has been disrupted. This pool of edited cells, generated by target-gene disruption via CRISPR/Cas9, offers a loss-of-function model for studying CCR5-dependent biological processes. Unlike clonal lines, the polyclonal format preserves heterogeneity in editing outcomes, enabling robust functional analysis while mitigating risks associated with clonal artifacts. The product is intended for advanced research applications in immunology, virology, and signal transduction.
The HAP1 cell line is a near-haploid human cell line derived from the chronic myeloid leukemia (CML) cell line KBM-7. Its near-haploid karyotype facilitates gene-editing studies and functional genomics screening. HAP1 cells retain intact GPCR signaling machinery and exhibit adherent growth properties suitable for diverse in vitro assays. This genetic background supports the analysis of receptor-mediated pathways without confounding effects from diploid gene redundancy, making it a valuable host for knockout modeling.
CCR5 encodes a G protein-coupled receptor that functions as a key chemokine receptor and a major HIV-1 co-receptor. The receptor is activated by chemokine ligands including CCL3 (MIP-1??), CCL4 (MIP-1??), CCL5 (RANTES), CCL8, and CCL11. Upon activation, CCR5 couples primarily to G??i proteins, leading to G?¦?-mediated signaling, adenylyl cyclase inhibition, and calcium mobilization. CCR5 interacts with CD4 and the HIV envelope glycoprotein gp120 to facilitate viral entry. Downstream signaling involves the PI3K-Akt pathway, MAPK/ERK cascade, and NF-??B activation, ultimately regulating cell migration and inflammatory responses. ??-arrestin-2 mediates receptor desensitization and additional signaling outputs.
Disruption of CCR5 in HAP1 cells abolishes its HIV-1 co-receptor function and impairs chemokine-induced chemotaxis. In the context of a haploid genetic background, this knockout model enables clear genotype-phenotype correlations. The polyclonal population provides a pool of diverse mutations, which is particularly useful for studying signaling pathways where partial or mosaic disruption may reveal dose-dependent effects. Researchers can co-culture these cells with relevant ligands or viral pseudotypes to dissect receptor requirements and downstream signaling intermediaries without interference from wild-type alleles.
This knockout model supports a broad range of experimental applications, including HIV pseudovirus entry assays to evaluate co-receptor dependence, Transwell migration assays to measure chemotactic responses, and calcium flux assays to monitor receptor-mediated intracellular signaling. Drug screening for CCR5 antagonists or inhibitors of HIV entry can be performed using these cells combined with flow cytometric analysis of surface marker expression. Validation of knockout status is readily achieved by Western blot for protein level, qPCR for transcript abundance, and flow cytometry for CCR5 surface expression. For further product details and technical support, please contact Ascent Research.
