The IL27 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This product features targeted disruption of the IL27 gene, generating a heterogeneous pool of edited cells suitable for studying IL27-dependent functions in intestinal epithelial biology. The polyclonal format maintains genetic diversity while eliminating IL27 expression, enabling robust loss-of-function analyses without the limitations of clonal selection. Researchers can interrogate IL27-mediated signaling pathways in an epithelial context that retains the capacity for enterocytic differentiation, providing a physiologically relevant platform for investigating cytokine crosstalk in the gastrointestinal tract.
HT29 cells, originally isolated from a primary colorectal adenocarcinoma, exhibit an epithelial morphology and can be induced to differentiate into polarized, enterocyte-like cells under appropriate culture conditions. This cell line is widely employed as an in vitro model for intestinal epithelial barrier function, colorectal cancer biology, and mucosal immune responses. The HT29 background expresses key cytokine receptors and signaling intermediates, making it a valuable host for exploring how tumor-derived factors such as IL27 modulate epithelial homeostasis, immune cell recruitment, and malignant progression within the tumor microenvironment.
IL27 is a heterodimeric cytokine composed of the EBI3 and p28 subunits. It signals through a receptor complex comprising IL27RA and gp130, leading to activation of JAK1 and JAK2 and subsequent phosphorylation of STAT1 and STAT3. These transcription factors drive expression of downstream targets including T-bet, IL-10, and SOCS3. IL27 exerts context-dependent immunomodulatory effects: it promotes Th1 differentiation and IFN-?? production while also inducing IL-10-mediated anti-inflammatory responses. Upstream regulators such as TLR ligands, interferons, CD40 ligation, and transcription factors NF-??B, IRF3, and IRF7 control IL27 expression. In epithelial cells, IL27 signaling intersects with pathways governing proliferation, apoptosis, and barrier integrity, highlighting its multifaceted role at the intestinal mucosa.
In the context of HT29 colorectal adenocarcinoma cells, IL27 knockout provides a powerful tool to dissect the cytokine??s contribution to tumor?Cimmune cell interactions and epithelial barrier regulation. IL27 produced by tumor cells or infiltrating immune cells can shape the inflammatory milieu, influencing cancer cell survival, migration, and cytokine secretion. This knockout model enables direct investigation of how loss of IL27 affects STAT1/STAT3 phosphorylation dynamics, downstream gene expression, and functional outcomes such as monolayer permeability. Given the association of IL27 with colorectal cancer and inflammatory bowel disease, the system is highly relevant for pre-clinical studies exploring IL27 as a therapeutic target or biomarker.
Typical applications include western blotting and RT-qPCR for confirming IL27 ablation, RNA-seq transcriptomic profiling to identify IL27-dependent gene networks, and flow cytometry or phospho-STAT analysis to monitor signaling alterations. Functional assays can assess cell proliferation, migration, invasion, and epithelial barrier integrity. Co-culture setups with immune cells allow modeling of epithelial?Cimmune crosstalk, while cytokine secretion assays quantify the impact on the local microenvironment. The polyclonal population is well-suited for drug screening campaigns targeting IL27 pathway components. For further information on this product, please contact Ascent Research.
