The GUF1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the SK-HEP-1 human liver adenocarcinoma cell line. This product features targeted disruption of the GUF1 gene, enabling loss-of-function studies in a heterogeneous cell pool that minimizes clonal bias. The polyclonal format provides a robust model for investigating mitochondrial translation without the limitations of single-cell clones.
SK-HEP-1 cells, originally isolated from the ascites of a liver adenocarcinoma patient, serve as a well-characterized in vitro model of hepatocellular carcinoma with epithelial morphology. They recapitulate key metabolic features of hepatic tumors and are extensively used in cancer metabolism and drug screening research. Their derivation from a metastatic site provides a relevant context for studying tumor cell energetics and mitochondrial adaptations.
GUF1 encodes a mitochondrial elongation factor essential for ribosome recycling after translation termination. It functions together with TUFM and TSFM elongation factors and interacts with mitochondrial ribosomal large and small subunit proteins and the release factor mtRF1a. GUF1 activity is regulated by mitochondrial ribosome stalling, amino acid deprivation, and the mitochondrial unfolded protein response (UPRmt), and it mediates the release of nascent peptides and ribosomal subunits from mt-mRNA. This recycling step is critical for maintaining mitochondrial protein synthesis efficiency and fidelity.
Knockout of GUF1 in SK-HEP-1 cells impairs mitochondrial translation, decreasing synthesis of mtDNA-encoded oxidative phosphorylation subunits such as MT-CO1 and MT-ND1. This defect leads to respiratory chain deficiency, reduced ATP production, and a compensatory increase in glycolysis, phenocopying features of mitochondrial diseases including lactic acidosis and combined oxidative phosphorylation deficiency. In the hepatocellular carcinoma context, the model enables exploration of how mitochondrial dysfunction alters tumor cell metabolism, survival, and stress responses.
The GUF1 knockout polyclonal cells are ideal for studying mitochondrial translation regulation and its impact on liver cancer metabolism. Representative applications include Western blotting for mitochondrial proteins (MT-CO1, MT-ND1), Seahorse metabolic flux analysis (oxygen consumption rate), ATP and lactate measurements, and flow cytometry for mitochondrial membrane potential using TMRM. The model also supports mitochondrial ribosomal profiling, immunofluorescence for morphology, co-immunoprecipitation of ribosome complexes, and drug screening for mitochondrial disorders. For additional information, please contact Ascent Research.
