The GUF1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HT29 colorectal adenocarcinoma cells. This product comprises a genetically diverse pool with targeted disruption of GUF1, a gene encoding a mitochondrial GTPase essential for mitochondrial translation elongation and ribosome recycling. The polyclonal format avoids clonal artifacts and enables robust study of GUF1 loss-of-function effects on mitochondrial protein synthesis and oxidative phosphorylation.
HT29 cells are an adherent epithelial line from a human colorectal adenocarcinoma, widely used as a model for intestinal biology and colorectal cancer research. They are particularly valued for investigating signaling, metabolism, and drug responses. Their well-characterized metabolic phenotype and mitochondrial activity make them an appropriate host to examine how GUF1 disruption influences mitochondrial function and tumor cell physiology.
GUF1 functions as a mitochondrial translation elongation factor with GTPase activity, facilitating efficient elongation and ribosomal recycling in the mitochondrial translation system. It interacts with mitochondrial elongation factor Tu (TUFM) and the mitoribosome, including subunits MRPL12 and MRPS proteins. GUF1 is critical for synthesis of mitochondrially encoded OXPHOS subunits such as MT-ND1, MT-CO1, and MT-ATP6. Its expression is regulated by the PGC-1??/NRF-1/TFAM transcriptional cascade, linking mitochondrial biogenesis to oxidative metabolism. Loss of GUF1 disrupts translation, leading to defective assembly of complexes I, IV, and V and impaired oxidative phosphorylation.
In HT29 cells, GUF1 knockout induces mitochondrial translation deficiency, impairing OXPHOS and promoting glycolytic shift. This phenotype recapitulates features of mitochondrial disorders like combined oxidative phosphorylation deficiency and lactic acidosis, while reflecting metabolic reprogramming seen in aggressive colorectal cancers. The model allows dissection of mitochondrial dysfunction effects on energy metabolism, redox homeostasis, and apoptosis in epithelial cancer cells, providing insights into metabolic vulnerabilities and stress responses in colorectal tumors.
Applications include Western blotting for OXPHOS subunits, RT-qPCR of mtDNA transcripts, Seahorse respirometry, flow cytometry for mitochondrial membrane potential and apoptosis, and drug sensitivity assays against OXPHOS. These cells enable clonogenic survival studies under metabolic stress and screening for mitochondrial function modulators. They also facilitate investigation of PGC-1??/NRF-1/TFAM signaling in colorectal cancer metabolism. For more information, please contact Ascent Research.
