HNF4A Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 143B osteosarcoma cell line, engineered to disrupt the HNF4A gene. The product provides a mixed population of cells with targeted gene disruption generated by CRISPR/Cas9, enabling loss-of-function studies of HNF4A without clonal selection. This polyclonal format maintains genetic heterogeneity while ensuring robust knockout efficiency across the population, suitable for experiments requiring a representative knockout model rather than an isogenic clone.
The 143B cell line is a highly tumorigenic and metastatic derivative of the HOS human osteosarcoma line, widely used as a model for bone cancer biology. Originating from a malignant bone tumor, 143B cells exhibit aggressive growth characteristics and a capacity for experimental metastasis, making them a relevant system for investigating molecular determinants of osteosarcoma progression. The absence of hepatocyte lineage-specific factors in this non-hepatic background allows researchers to study HNF4A??s transcriptional functions without confounding hepatic differentiation programs.
HNF4A encodes a nuclear receptor transcription factor that acts as a master regulator of hepatocyte differentiation and metabolic homeostasis. It transcriptionally activates a range of downstream target genes including APOA1, CYP7A1, PCK1, and HNF1A, thereby controlling pathways such as cholesterol efflux, bile acid synthesis, gluconeogenesis, and hepatocyte maintenance. HNF4A activity is modulated by upstream regulators like HNF1A, FOXA2, and WNT/??-catenin signaling, and it forms heterodimers with RXRA and complexes with coactivators such as PPARGC1A or corepressors like NCOR2. Disruption of HNF4A in this model ablates its direct transcriptional control over metabolic gene networks, providing a clean system to dissect HNF4A-dependent regulation.
In the 143B osteosarcoma context, HNF4A knockout permits investigation of its role outside the liver, where its expression may contribute to metabolic reprogramming in cancer cells. Osteosarcoma cells often rewire metabolic pathways, and HNF4A could influence lipid and glucose utilization. This model enables deconvolution of HNF4A-mediated transcriptional programs independently of hepatic lineage-specific cofactors, facilitating studies on its ectopic functions in tumor metabolism. Moreover, it allows assessment of HNF4A??s interaction with oncogenic signaling pathways such as HGF/MET or WNT/??-catenin in a malignant bone tumor environment.
This polyclonal knockout population is valuable for functional genomics of metabolic transcription factors, drug metabolism studies, and exploring HNF4A??s role in non-hepatic cancers. Typical assays include Western blotting to confirm loss of HNF4A protein, RT-qPCR to measure downregulation of targets like APOA1 and ALB, RNA-seq for global transcriptome profiling, ChIP-qPCR to evaluate occupancy at genomic binding sites, luciferase reporter assays for transcriptional activity, metabolic flux analyses, and drug sensitivity testing. The model supports CRISPR knockout validation and can serve as a starting point for hepatocyte differentiation studies where HNF4A??s function is investigated. For further information, please contact Ascent Research.
