The CCR7 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line, engineered to disrupt the CCR7 gene. This heterogeneous pool of gene-edited cells offers a practical loss-of-function model for investigating the roles of CCR7 in chemokine receptor signaling and immune cell trafficking, without requiring isolation of single-cell clones.
The HAP1 cell line is a near-haploid, adherent fibroblast-like line originating from the KBM-7 chronic myeloid leukemia (CML) line, which was derived from a 39-year-old male with CML in blast crisis. Its near-haploid genome facilitates straightforward genetic analysis, as disruption of a single allele can result in functional protein loss. Despite its transformed origin, HAP1 retains hematopoietic features and has become a standard model for studying signal transduction, migration, and cancer biology.
CCR7 encodes a G protein-coupled chemokine receptor that specifically recognizes the homeostatic chemokines CCL19 and CCL21. Upon ligand binding, the receptor activates heterotrimeric G??i proteins, triggering downstream signaling cascades that involve PI3K/AKT, MAPK/ERK, and Rho GTPase modules. Key downstream targets include AKT, ERK1/2, p38 MAPK, and the NF-??B p65 subunit, which become phosphorylated, while the small GTPases Rac1 and RhoA govern actin polymerization and cell polarization for migration. CCR7 signaling is regulated by GRK-mediated phosphorylation and ??-arrestin-2 recruitment, and it functionally interacts with other chemokine receptors such as CXCR4 and CCR5. Inflammatory stimuli like TNF-?? can also modulate CCR7 expression.
In the HAP1 context, CCR7 knockout provides a unique system to dissect the signaling mechanisms that control hematopoietic cell migration and metastasis. The leukemic background of the host cells mirrors aspects of diseases such as chronic lymphocytic leukemia, multiple sclerosis, and rheumatoid arthritis, where CCR7 has been implicated. Disruption of CCR7 in these cells enables examination of how loss of this receptor impacts downstream effectors, cytoskeletal dynamics, and cell motility, offering insights into potential therapeutic interventions targeting the chemokine signaling axis.
This polyclonal knockout cell pool is suited for a wide range of functional assays, including Transwell migration assays, flow cytometric measurement of CCR7 surface expression, western blotting for phospho-AKT and phospho-ERK1/2, and adhesion assays assessing LFA-1 integrin activation. It serves as a valuable resource for cancer metastasis research, immunology, T cell biology, and drug discovery programs aimed at chemokine receptor pathways. By providing a straightforward loss-of-function system, these cells enable detailed study of CCR7-dependent signaling in disease. For more information, please contact Ascent Research.
