The HNF4A Knockout 786-O Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human clear cell renal cell carcinoma line. This product provides a heterogeneous pool of cells with targeted disruption of HNF4A, avoiding clonal artifacts while enabling robust loss-of-function studies in renal cancer research.
The 786-O host cell line was originally established from a primary clear cell renal cell carcinoma and is characterized by a VHL mutation that leads to constitutive HIF activation. It displays adherent epithelial morphology and is extensively used as a model for ccRCC to study hypoxia-driven tumorigenesis, metabolic reprogramming, and drug sensitivity. This genetic background offers a defined platform for interrogating HNF4A??s contribution to renal cancer biology.
HNF4A functions as a ligand-independent nuclear receptor that binds direct repeat DNA elements as a homodimer to transcriptionally regulate genes pivotal to hepatocyte and renal epithelial function. Its activity is governed by upstream regulators such as HNF1A, PPARA, HIF1A, glucocorticoids, insulin, and PGC1A, and by metabolic cues like fatty acids. Key downstream effectors include CYP3A4, PCK1, SLC2A2, APOB, ALB, and various solute carriers and fatty acid binding proteins. HNF4A interacts with coactivators PGC1A and SRC1, corepressor NCOR1, and transcriptional partners HNF1A, FOXA2, and SMARCD1. Disruption of HNF4A compromises the HNF4A/HNF1A regulatory loop and PGC1A coactivation, leading to diminished expression of genes involved in gluconeogenesis, drug metabolism, and epithelial transport.
In the 786-O renal carcinoma setting, loss of HNF4A disrupts the epithelial differentiation program and metabolic enzyme networks essential for ccRCC survival. VHL-mutated tumors rely on metabolic adaptation, and HNF4A knockout enables dissection of the transcription factor??s role in fatty acid metabolism and solute carrier expression, independently of the HIF-driven axis. This model provides insight into how HNF4A coordinates with hypoxia signaling to maintain the metabolic and epithelial characteristics of renal cancer cells.
Applications span metabolic flux analysis, RNA-seq, ChIP-seq, and RT-qPCR for characterizing HNF4A targets and metabolic shifts. Researchers can perform western blotting, reporter assays, migration and invasion assays, drug sensitivity panels, and immunofluorescence to evaluate HNF4A-dependent phenotypes. This knockout cell pool is especially suited for studies on metabolic reprogramming, tumor biology, and drug transporter regulation in renal cell carcinoma. Contact Ascent Research for further details.
