The AAMDC Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Huh-7 human hepatocellular carcinoma line, engineered to disrupt AAMDC expression. This heterogeneous loss-of-function model enables investigation of AAMDC-dependent processes in a liver epithelial background without single-cell cloning, capturing a broad spectrum of gene-editing events for population-level functional studies.
The parental Huh-7 cell line originates from a well-differentiated hepatocellular carcinoma and serves as a widely used model of human hepatocyte function. Huh-7 cells display epithelial morphology and retain key metabolic activities, including lipoprotein synthesis, drug-metabolizing enzyme expression, and insulin-responsive signaling. They are extensively employed to study hepatic lipid metabolism, mitochondrial bioenergetics, and the pathogenesis of NAFLD and viral hepatitis, providing a physiologically relevant platform for investigating mitochondrial protein function in liver disease.
AAMDC encodes a conserved mitochondrial matrix protein that directly interacts with LRPPRC, a regulator of mitochondrial mRNA stability and translation. It operates downstream of insulin signaling and is transcriptionally regulated by PPARA, PPARG, and SREBF1. Disruption of AAMDC diminishes expression of mitochondrial respiratory chain subunits and fatty acid oxidation enzymes such as CPT1 and ACOX1, leading to compromised oxidative phosphorylation, reduced ??-oxidation, and elevated ROS levels, thereby destabilizing hepatocellular lipid homeostasis and bioenergetic capacity.
In the Huh-7 hepatocellular carcinoma context, AAMDC knockout creates a relevant model for studying mitochondrial dysfunction and hepatic steatosis. Loss of AAMDC-dependent regulation of fatty acid oxidation and respiratory efficiency recapitulates key metabolic perturbations observed in NAFLD and obesity-related liver disease. This polyclonal knockout population enables dissection of how mitochondrial defects drive excessive lipid storage, impair insulin sensitivity, and trigger pro-apoptotic signaling, providing insight into early molecular events of metabolic liver disease progression.
These AAMDC knockout Huh-7 polyclonal cells are suited for multifaceted experimental workflows. Lipid accumulation can be assessed by Oil Red O staining, while mitochondrial function is profiled via Seahorse extracellular flux analysis. Mitochondrial membrane potential and apoptosis are monitored by flow cytometry. Expression changes in PPAR targets, respiratory chain subunits, and fatty acid oxidation enzymes are analyzed by RT-qPCR and Western blotting. The polyclonal population is ideal for high-content screening for compounds that correct lipid dysregulation or restore mitochondrial respiration in NAFLD models. For support, contact Ascent Research.
