The IL27 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HCT 116 human colorectal carcinoma cells in which the IL27 gene has been disrupted to ablate IL-27 cytokine expression. This polyclonal pool contains a heterogeneous set of gene-edited cells, providing a robust loss-of-function model that avoids the limitations of single-clone selection. The target gene disruption is achieved using CRISPR/Cas9, resulting in elimination of endogenous IL-27 production.
The HCT 116 cell line is a widely used epithelial model of colon carcinoma, derived from a male patient. It exhibits microsatellite instability (MSI-high) due to MLH1 deficiency and carries activating mutations in KRAS (G13D) and CTNNB1 (??-catenin). These genetic features drive constitutive Wnt signaling and a hypermutator phenotype, making it a valuable system for studying colorectal cancer biology, drug responses, and tumor immunology.
IL-27 is a heterodimeric cytokine comprising p28 and EBI3 subunits that signals through the IL27RA/gp130 receptor complex. This interaction activates receptor-associated JAK1, JAK2, and TYK2 kinases, leading to phosphorylation and nuclear translocation of STAT1 and STAT3. Downstream transcriptional targets include T-bet, IL10, IL12RB2, and SOCS1/3, which mediate IL-27??s immunomodulatory functions. Upstream regulators such as NF-??B, IRF3, and IRF7 control IL27 expression in response to TLR ligands, CD40 engagement, and IFN-??. In the knockout cells, this signaling pathway is disrupted, eliminating the cellular effects of IL-27.
In the HCT 116 context, loss of IL-27 enables dissection of its role in tumor-intrinsic processes and microenvironmental interactions. Given the cell line??s oncogenic ??-catenin and KRAS signaling, IL-27 depletion may alter proliferation, apoptosis, and inflammatory gene expression. These polyclonal knockout cells are thus instrumental for investigating cytokine-mediated crosstalk in colon cancer, as well as for evaluating IL-27 as a therapeutic target or biomarker.
Applications include tumor immunology, cytokine signaling, cancer microenvironment studies, and drug target validation. Typical assays encompass western blotting and ELISA for IL-27, RT-qPCR for downstream targets, phospho-STAT flow cytometry, proliferation (MTT/BrdU) and apoptosis (Annexin V) assays, immune cell co-cultures, and RNA-seq. For further inquiries, contact Ascent Research.
