The GTPBP10 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in the HAP1 near-haploid human cell line, engineered for loss-of-function analysis of the GTPBP10 gene. This product provides a targeted disruption of GTPBP10, encoding a mitochondrial GTPase essential for assembly of the small subunit of the mitochondrial ribosome. It serves as a robust model system to dissect mitochondrial translation and ribosome biogenesis pathways.
HAP1 cells are a near-haploid, adherent cell line derived from a male chronic myeloid leukemia patient harboring the Philadelphia chromosome. With a haploid karyotype for most chromosomes, a single genetic disruption can produce a full knockout phenotype, making HAP1 a preferred platform for functional genomic screens. The cells express hematopoietic lineage markers and are adapted to standard adherent culture, ensuring compatibility with routine cell biology techniques.
GTPBP10 functions as a mitochondrial GTPase that facilitates the maturation of the small subunit (28S) of the mitochondrial ribosome. It is downstream of mitochondrial import machinery and TFAM, and is responsive to cellular energy status. GTPBP10 directly interacts with MRPS proteins, mitoribosome assembly factors, mitochondrial rRNA, and GTP to drive ribosomal subunit assembly. This process is critical for mitochondrial translation, promoting synthesis of mtDNA-encoded subunits of oxidative phosphorylation complexes, including MT-CO1 and MT-CYB. Disruption of GTPBP10 thus impairs mitochondrial gene expression, linking its function to mitochondrial disorders and potential roles in neurodegenerative diseases.
Exploiting the haploid nature of HAP1, CRISPR/Cas9-mediated knockout of GTPBP10 yields a clean loss-of-function phenotype unconfounded by a second allele, making this polyclonal population highly effective for studying mitochondrial translation defects. Impaired ribosome assembly can be assessed by sucrose gradient fractionation profiling, while mitochondrial protein synthesis rates are quantifiable via radioactive pulse-labeling. The model is particularly valuable for modeling mitochondrial dysfunction caused by defective mitoribosome biogenesis and for investigating how these defects contribute to cellular pathology.
The GTPBP10 Knockout HAP1 Polyclonal Cells support diverse experimental applications, including high-content imaging of mitochondrial morphology, growth assays on galactose medium to evaluate oxidative phosphorylation reliance, and RT-qPCR for mitochondrial transcript levels. Western blotting for mitochondrial-encoded subunits such as MT-CO1 provides confirmation of downstream effects on respiratory chain expression. This knockout model is also suited for drug screening campaigns targeting mitochondrial diseases and functional genomics studies of mitochondrial ribosome assembly. For further information, please contact Ascent Research.
