The CCL25 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which CCL25 gene disruption creates a loss-of-function model. This polyclonal product provides a heterogeneous pool of edited cells suitable for functional studies without clonal selection artifacts. The CRISPR/Cas9-mediated gene disruption efficiently abrogates CCL25 expression, enabling functional genomics and pathway analysis in a human cellular context.
The parental HAP1 cell line is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia. Its near-haploid karyotype simplifies genetic manipulation, as only a single allele requires targeting to generate complete gene disruptions. HAP1 cells exhibit robust growth and are amenable to various assays including chemotaxis, signaling analysis, and cytokine quantification. Their human origin ensures translational relevance, and the leukemic background provides context for studying chemokine-mediated processes in hematological malignancies.
CCL25 is a chemokine that serves as a chemoattractant for CCR9-expressing cells, primarily orchestrating T lymphocyte homing to the small intestine. Upon binding CCR9, CCL25 triggers G-protein-coupled signaling, leading to calcium mobilization and activation of PI3K/Akt and MAPK pathways. This cascade promotes integrin-dependent adhesion and directed migration. CCL25 expression is induced by pro-inflammatory cytokines including TNF-alpha, IL-1beta, and TGF-beta, acting through NF-??B. The CCL25/CCR9 axis interacts with glycosaminoglycans and matrix metalloproteinases, fine-tuning leukocyte trafficking. Key downstream effectors include PI3K, AKT, MAPK, PLC, and G proteins.
In the HAP1 background, CCL25 knockout eliminates autocrine or paracrine chemokine signals, enabling dissection of CCL25-dependent signaling in a simplified genetic system. Researchers can probe CCR9-mediated responses to exogenous CCL25 in migration and invasion assays, or study how loss of CCL25 impacts baseline signaling and gene expression. The near-haploid nature facilitates synthetic lethality screens and genetic interaction studies aimed at identifying CCL25 pathway modulators in leukemia or inflammation.
Key applications include Transwell chemotaxis and calcium flux assays to assess signaling dynamics, Western blotting for phospho-AKT and ERK, and ELISA for CCL25 secretion. RT-qPCR and flow cytometry allow measurement of CCL25 transcript and CCR9 surface expression, respectively. These cells support drug target validation for the CCL25/CCR9 axis, inflammatory bowel disease models, and mucosal immunity research. For further information, please contact Ascent Research.
