The IL1RN Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line, with targeted disruption of the IL1RN gene. This heterogeneous pool serves as a loss-of-function model for studying interleukin-1 receptor antagonist (IL-1ra) biology, free from clonal selection biases. CRISPR/Cas9-mediated gene disruption yields a mixed population of cells with targeted modifications at the IL1RN locus.
The HT29 cell line originates from a human colorectal adenocarcinoma and serves as a prominent intestinal epithelial model. These cells form differentiated polarized monolayers, produce mucins, and exhibit characteristics of absorptive enterocytes, making them widely used for studies of gut barrier function, drug transport, and colorectal cancer pathogenesis. The colorectal epithelial context is especially pertinent for examining inflammatory processes regulated by IL-1 signaling.
IL1RN encodes IL-1ra, a secreted antagonist that competes with IL-1?? and IL-1?? for binding to IL-1R1. IL-1ra occupies the receptor but fails to recruit IL-1RAcP, thereby preventing assembly of the signaling complex required for MyD88-dependent recruitment of IRAK1, IRAK4, and TRAF6. This halts activation of NF-??B and MAPK (ERK, JNK, p38) pathways, leading to suppression of downstream pro-inflammatory effectors including IL-6, IL-8, TNF-??, and COX-2. IL1RN transcription is itself activated by NF-??B and AP-1 in response to IL-1??, TNF-??, and LPS, forming an autoregulatory circuit.
Knockout of IL1RN in HT29 cells ablates the endogenous IL-1 signaling brake, rendering the cells hyper-responsive to IL-1?? and other inflammatory stimuli. These polyclonal knockout cells are expected to show enhanced NF-??B and MAPK phosphorylation, elevated IL-6 and IL-8 secretion, and broad transcriptional reprogramming upon cytokine challenge, mirroring aspects of intestinal inflammation and tumor microenvironment signaling observed in vivo in conditions such as inflammatory bowel disease and colorectal cancer.
Researchers can leverage this model to screen IL-1 pathway inhibitors, investigate IL-1-driven epithelial barrier disruption using TEER assays, or perform cytokine profiling via ELISA and RNA-seq. The polyclonal knockout pool is also suitable for NF-??B reporter assays, co-culture experiments, and mechanistic dissection of IL-1 feedback loops. For further technical specifications and batch-specific data, please contact Ascent Research.
