The HNF4A Knockout DLD-1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal population of Homo sapiens DLD-1 colorectal adenocarcinoma epithelial cells with targeted disruption of the HNF4A gene. This polyclonal knockout pool offers a heterogeneous loss-of-function model, avoiding clonal selection artifacts and enabling robust assessment of HNF4A-dependent phenotypes. Cells are supplied ready-to-use with knockout confirmed by genomic screening, allowing immediate culture for investigation of gene function without single-cell clonal variation.
The host DLD-1 cell line, derived from a male Dukes?? type C colorectal adenocarcinoma patient, exhibits epithelial morphology and metastatic potential. It harbors oncogenic mutations in APC, KRAS, TP53, PIK3CA, and SMAD4, reflecting the genetic profile of aggressive colorectal cancer and making it an established model for colon carcinogenesis, invasion, and therapy response. DLD-1 cells form cohesive monolayers expressing key cell?Ccell junction components, providing a physiologically relevant context for HNF4A-mediated intestinal epithelial regulation studies.
HNF4A is a nuclear receptor transcription factor that homodimerizes and drives hepatocyte and intestinal epithelial differentiation, metabolic regulation, and tight junction integrity. It activates CDH1 and CLDN1, and is controlled by upstream regulators HNF1A, FOXA2, and PPARGC1A, engaging coactivators like NCOA1 and EP300. Key downstream effectors include APOA1, CYP7A1, CYP3A4, and ABCB1, connecting HNF4A to lipid metabolism, drug disposition, and epithelial barrier function.
In DLD-1 cells, HNF4A knockout disrupts enterocyte differentiation and junctional assembly, compromising barrier integrity and altering metabolic signatures. This model is critical for investigating how HNF4A loss cooperates with APC, KRAS, and TP53 mutations to enhance proliferation, migration, and metabolic reprogramming in colorectal cancer. It also enables MODY1 metabolic defect modeling and evaluation of HNF4A tumor-suppressive or oncogenic roles. The polyclonal format maintains genetic heterogeneity, better mirroring clinical tumor populations than monoclonal isolates.
Researchers can apply this knockout pool for western blotting of HNF4A, CDH1, and CLDN1; RT-qPCR of APOA1 and CYP3A4; immunofluorescence staining of ZO-1 and occludin; and TEER-based barrier integrity measurements. RNA-seq can define HNF4A-dependent transcriptomes, and metabolic assays for glucose uptake and lipid accumulation reveal functional consequences. ChIP and co-immunoprecipitation studies map DNA-binding and interactions with NCOR1 or PPARGC1A. The model supports drug metabolism research, inflammatory bowel disease investigation, and therapeutic targeting studies. For technical details, contact Ascent Research.
