The CCR1 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human CCR1 gene within the near-haploid HAP1 cell line. This pool of CRISPR-engineered cells carries heterogeneous loss-of-function alleles, providing a versatile and robust model for studying CCR1-dependent biological processes. The polyclonal format ensures a broad representation of genetic disruptions, enabling statistically rigorous comparisons and phenotypic screening without the constraints of clonal variability.
The HAP1 cell line is derived from KBM-7 chronic myeloid leukemia cells and exhibits a near-haploid karyotype, making it an exceptional host for genetic modification and functional genomics. Its haploid nature simplifies gene disruption studies, as a single targeted allele typically yields a complete loss of function. HAP1 cells maintain stable growth characteristics, are amenable to high-throughput assays, and are widely used as a platform for chemical and genetic screening, providing a clean cellular background for investigating signaling pathways and drug targets.
CCR1 encodes a G protein-coupled receptor that serves as the primary receptor for CC chemokines including CCL3 (MIP-1??), CCL5 (RANTES), and CCL7 (MCP-3). Ligand engagement activates G??i heterotrimeric G proteins, leading to the release of G?¦? subunits, which stimulate PLC??, generate IP3, and mobilize intracellular calcium. This cascade further triggers the activation of PI3K-AKT and the Ras-Raf-MEK-ERK MAP kinase pathway, as well as p38 and JNK, ultimately promoting the activation of transcription factors such as NF-??B and AP-1. These transcriptional events drive the expression of genes involved in integrin activation, cytokine production, and leukocyte chemotaxis. Additionally, CCR1 interacts with GRK2/3 and ??-arrestin1/2 for receptor desensitization and internalization, and can heterodimerize with CCR5, modulating signaling specificity.
In HAP1 cells, disruption of CCR1 abrogates chemokine-induced calcium flux, blocks PI3K/AKT and ERK phosphorylation, and eliminates NF-??B-mediated transcriptional responses, thereby creating a clean loss-of-function background. The polyclonal nature of this knockout pool reduces the risk of clonal artifacts and ensures that the observed phenotypes are truly representative of CCR1 ablation. This makes it particularly suitable for high-throughput applications where consistent and reproducible results are critical, such as compound library screening for receptor antagonists or synthetic lethal interactions.
Researchers can employ these polyclonal knockout cells in a variety of downstream assays, including chemotaxis assays (Boyden chamber), real-time calcium flux measurements, Western blotting for phospho-ERK and phospho-AKT, flow cytometric assessment of receptor surface expression, and RT-qPCR or ELISA to quantify alterations in cytokine and chemokine production. The cells are especially valuable for high-throughput chemical screens designed to identify novel CCR1 antagonists and for functional validation of the receptor as a therapeutic target in inflammatory disorders, autoimmunity, and cancer. For comprehensive technical support and ordering information, please contact Ascent Research.
