The IL17RA Knockout Caco-2 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human IL17RA gene in the Caco-2 intestinal epithelial background. This product consists of a heterogeneous pool of cells carrying various disruptions at the IL17RA locus, establishing a robust loss-of-function model that avoids clonal selection bias. The polyclonal format is particularly suited for functional genomics studies where population-level genetic perturbation better recapitulates tissue heterogeneity and reduces artifacts associated with single-cell expansion.
The parental Caco-2 line, derived from a human colorectal adenocarcinoma, is a well-established in vitro model of the intestinal epithelial barrier. Upon differentiation, these cells form polarized monolayers with functional tight junctions, apical microvilli, and appropriate transporter expression, making them ideal for studying barrier integrity, drug absorption, and mucosal immune responses. This epithelial origin makes the knockout derivative directly relevant to intestinal inflammation research and colorectal cancer biology.
IL17RA encodes the high-affinity receptor for the pro-inflammatory cytokines IL-17A and IL-17F. Ligand binding induces receptor complex formation with IL-17RC, recruiting the adaptor Act1 (TRAF3IP2) and the E3 ubiquitin ligase TRAF6. Downstream, TRAF6-mediated ubiquitination activates TAK1 and the IKK complex, resulting in NF-??B nuclear translocation and activation of the MAPK/AP-1 and C/EBP pathways. This signaling cascade drives transcription of inflammatory mediators such as IL-6, IL-8, the chemokine CXCL1, and antimicrobial defensins. CRISPR-mediated disruption of IL17RA in these polyclonal cells eliminates ligand-dependent signaling, preventing Act1/TRAF6 scaffolding and subsequent NF-??B, MAPK, and C/EBP activation.
In Caco-2 monolayers, IL17RA signaling governs epithelial inflammatory responses, chemokine secretion, and modulation of tight junction proteins. This knockout model therefore permits precise investigation of epithelial-specific contributions to IL-17-driven pathology, independent of immune cell crosstalk. The model is highly relevant for dissecting mechanisms of inflammatory bowel disease, where IL-17 promotes mucosal barrier dysfunction, and for exploring the tumor-promoting inflammatory microenvironment in colorectal cancer. The polyclonal nature ensures that the observed phenotypes reflect the average behavior of a genetically diverse cell population, enhancing translational relevance.
Typical applications include cytokine and chemokine profiling by RT-qPCR or ELISA (e.g., IL-6, IL-8, CXCL1), analysis of NF-??B pathway activation via phospho-p65 western blotting or immunofluorescence detection of p65 nuclear localization, and co-immunoprecipitation of IL17RA with Act1 to verify disrupted receptor complexes. Flow cytometry for surface IL17RA confirms target protein loss, while transepithelial electrical resistance (TEER) measurements evaluate barrier function under inflammatory challenge. These assays support research in colitis models, colorectal cancer inflammation, and high-throughput screening of anti-IL-17 therapeutics. For further inquiries, please contact Ascent Research.
