The IL3 Knockout HT29 Polyclonal Cells product comprises a polyclonal population of HT29 colorectal adenocarcinoma cells with CRISPR/Cas9-mediated disruption of the IL3 gene, creating a reliable loss-of-function model for interleukin-3 research. The polyclonal nature maintains genetic heterogeneity while abolishing functional IL3 protein, supporting robust bulk assays and avoiding clonal selection bias. This format is ideal for experiments requiring a representative knockout profile in a defined genetic background.
The HT29 cell line originates from a primary colorectal adenocarcinoma of a 44-year-old female and exhibits an adherent epithelial morphology with mucin production. It harbors mutant p53, high microsatellite instability (MSI-H), and the BRAF V600E mutation, driving constitutive MAPK pathway activity. These characteristics render HT29 a widely used model for colorectal cancer research, particularly for mismatch repair-deficient tumors and drug permeability studies.
IL3 encodes a hematopoietic growth factor that signals through the IL3RA/CSF2RB receptor complex, activating JAK2, which phosphorylates STAT5, and subsequently triggering MAPK/ERK and PI3K-AKT cascades. Upstream regulators include NFAT, NF-??B, AP-1, and pro-inflammatory cytokines; downstream targets encompass BCL2, BCL-xL, MCL1, MYC, CCND1, and PIM1, with feedback inhibition by CISH and SOCS proteins. In the tumor microenvironment, autocrine/paracrine IL3 signaling can promote cell proliferation, survival, and inflammation.
In HT29 cells, IL3 knockout likely disrupts autocrine or paracrine signaling loops, reducing JAK/STAT, MAPK, and PI3K/AKT pathway activation. This attenuation may impair tumor cell survival and proliferation, particularly in the presence of oncogenic BRAF V600E. The model enables dissection of the interplay between constitutive BRAF?CMAPK signaling and cytokine-driven inputs, shedding light on inflammatory mechanisms in MSI-H colorectal cancer progression and chemoresistance.
These polyclonal knockout cells are suitable for diverse functional studies, including Western blotting, RT-qPCR, proliferation (MTT/BrdU), apoptosis (Annexin V/PI), colony formation, and migration/invasion assays. Additional applications comprise ELISA-based cytokine profiling, RNA-seq transcriptomics, and drug sensitivity testing to evaluate IL3-dependent therapeutic responses. This model supports colorectal cancer research, cytokine signaling investigation, and drug target validation. For more information, contact Ascent Research.
