The ADIRF Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt ADIRF gene expression in human HT29 colorectal adenocarcinoma cells. This loss-of-function model supports functional studies of ADIRF??s roles in regulating adipogenesis, cell differentiation, and tumor suppression. The polyclonal format retains genetic diversity, minimizing clonal bias and providing a physiologically relevant tool for examining ADIRF-dependent pathways. The cells are supplied as a ready-to-use culture for downstream molecular and cellular assays.
HT29 is a human colorectal adenocarcinoma cell line derived from a primary tumor of a 44-year-old female. These cells display an intestinal epithelial phenotype, including mucin production and enterocytic differentiation capacity. Notably, HT29 carries a mutant TP53 gene, impairing DNA damage responses and enhancing survival. This background creates a permissive context to investigate additional tumor suppressors like ADIRF, making the knockout model particularly suitable for studying colorectal cancer progression mechanisms.
ADIRF acts downstream of the adipogenic transcription factors PPAR-gamma and C/EBP-alpha, integrating TGF-beta and p53 signaling. Mechanistically, ADIRF interacts with C/EBP transcription factors and co-regulators to promote p21/CDKN1A and BAX expression, while suppressing anti-apoptotic BCL2 family members. Its loss thus disrupts cell cycle arrest and apoptosis programs. DNA methylation at the ADIRF locus serves as an upstream repressive mechanism, linking epigenetic regulation to metabolic and oncogenic outcomes. The polyclonal knockout cells allow dissection of ADIRF??s coordination between adipogenesis and tumor suppression.
In HT29 cells, ADIRF knockout accelerates malignant phenotypes by unleashing unchecked proliferation and apoptosis resistance, synergizing with endogenous mutant p53. This model is valuable for exploring crosstalk between obesity-driven adipogenic signaling and colorectal cancer, as ADIRF bridges metabolic and tumor-suppressive pathways. Researchers can assess how ADIRF loss alters differentiation, mucin production, and invasive behavior. The polyclonal composition further enables analysis of heterogeneous responses that mirror tumor heterogeneity in vivo.
Key applications include functional analysis of ADIRF as a tumor suppressor, investigation of adipogenesis-colorectal cancer crosstalk, and drug screening for ADIRF reactivation. Compatible assays comprise Western blotting, RT-qPCR for p21 and BAX, MTT and colony formation assays, Annexin V/PI apoptosis detection, flow cytometric cell cycle analysis, RNA-seq profiling, and xenograft tumor models. These approaches facilitate comprehensive dissection of ADIRF-mediated signaling networks. For further details, contact Ascent Research.
