The GTPBP10 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human SK-HEP-1 liver adenocarcinoma line, engineered to disrupt the gene encoding the essential mitochondrial GTPase GTPBP10. This polyclonal knockout product provides a heterogeneous pool of cells, avoiding the biases of clonal selection, and serves as a robust loss-of-function model for investigating GTPBP10??s role in mitochondrial ribosome biogenesis and translation.
The SK-HEP-1 host cell line is an epithelial line originally isolated from the ascites of a patient with liver adenocarcinoma. It is widely employed in hepatocellular carcinoma research for studying tumor cell metabolism, metastatic progression, and drug metabolism. These cells retain key hepatic cancer characteristics, establishing a pathophysiologically relevant system to examine the consequences of mitochondrial dysfunction in liver malignancies.
GTPBP10 encodes a mitochondrial GTPase that physically interacts with the mitoribosome, driving its assembly and enabling the translation of the 13 mtDNA-encoded subunits of oxidative phosphorylation complexes. Its expression is regulated by upstream transcription factors NRF1 and TFAM, which coordinate mitochondrial biogenesis, and it responds to mitochondrial stress signals. The protein interacts with mitoribosome assembly factors such as MTG1 and MALSU1, as well as mitochondrial ribosomal proteins. Downstream, GTPBP10 promotes synthesis of essential OXPHOS components like MT-CO1 and MT-ND1. Consequently, disruption of GTPBP10 impairs mitochondrial translation, leading to respiratory chain deficiencies and compromised cellular bioenergetics.
The GTPBP10 knockout in SK-HEP-1 cells is particularly informative because hepatocellular carcinoma frequently exhibits metabolic rewiring with increased reliance on mitochondrial respiration. This polyclonal knockout model permits exploration of how mitoribosome biogenesis influences tumor cell fitness, survival under metabolic stress, and sensitivity to mitochondrial inhibitors. Population-level assays avoid clonal adaptation, better reflecting the heterogeneous behavior of tumor cells.
Applications include western blotting for OXPHOS subunits, RT-qPCR of mitochondrial transcripts, Seahorse respirometry, puromycin labeling of mitochondrial translation, clonogenic survival, and apoptosis assays. The cells support mitochondrial ribosome profiling and can be used to screen for inhibitors of mitochondrial protein synthesis or to characterize metabolic reprogramming upon GTPBP10 loss. For more information, contact Ascent Research.
