Description

The ACSL5 Knockout HuH-7 Cell Line is a CRISPR/Cas9-edited knockout cell line that disrupts the ACSL5 gene in HuH-7 human hepatocellular carcinoma cells. This loss-of-function model enables stable and reproducible ablation of ACSL5 expression, facilitating detailed investigation of fatty acid metabolism and lipid signaling pathways in a hepatic context.

HuH-7 cells originate from a well-differentiated hepatocellular carcinoma of a 57-year-old male and retain liver-specific functions, making them an ideal model for liver cancer, hepatocyte metabolism, and viral hepatitis research. Their metabolic competence supports physiologically relevant studies of hepatic lipid handling, providing a robust background for interrogating ACSL5 function.

ACSL5 encodes a long-chain fatty acyl-CoA synthetase that catalyzes the thioesterification of fatty acids with coenzyme A, a critical activation step. It is transcriptionally regulated by PPAR??, SREBP-1c, ChREBP, and LXR, and responds to insulin, glucose, and polyunsaturated fatty acids. The resulting acyl-CoAs are directed to CPT1A for ??-oxidation, DGAT1/2 for triglyceride synthesis, and phospholipid biosynthetic enzymes, also contributing to pro-inflammatory eicosanoids. ACSL5 interacts with fatty acid transporters FATP2/4, lipid droplet proteins, and MBOAT7, and operates upstream of PPAR?? transcriptional programs controlling lipid catabolism and energy homeostasis.

In HuH-7 cells, ACSL5 disruption impairs fatty acid activation, reducing acyl-CoA pools. This compromises triglyceride and phospholipid synthesis, attenuates fatty acid oxidation, and dampens PPAR?? signaling. Consequently, the knockout model often exhibits reduced lipid accumulation, altered membrane composition, and modified cell proliferation??phenotypes relevant to hepatic steatosis, hepatocellular carcinoma, and metabolic syndrome. Thus, it serves as a powerful tool for dissecting lipid metabolic dysfunction.

This cell line supports diverse applications: mechanistic studies of lipid metabolism in liver cancer, lipid droplet dynamics, and drug screening for NAFLD. It is suitable for Western blotting, RT-qPCR, fatty acid oxidation assays, triglyceride quantification, Oil Red O staining, lipidomics, Seahorse respiration analysis, and flow cytometry. The model also enables transcriptomic and interactome studies to elucidate ACSL5-dependent networks. For further details, please contact Ascent Research.