The GON7 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the GON7 gene. This product employs non-clonal, polyclonal knockout cells that harbor heterogeneous disruptions in GON7, generated via CRISPR/Cas9-mediated gene disruption, avoiding isolation artifacts. As a knockout model, it abrogates GON7 protein expression, providing a robust system to dissect GON7-dependent cellular mechanisms.
HEK293T cells, derived from human embryonic kidney epithelia, are stably transfected with the SV40 large T antigen, enabling episomal replication of plasmids with SV40 ori. This characteristic makes the line exceptionally suitable for viral packaging and high-level recombinant protein expression. The well-annotated genome, rapid growth, and high transfectability of HEK293T provide a versatile background for gene editing applications, facilitating both mechanistic and screening studies.
The GON7 protein is an integral structural subunit of the KEOPS complex, which also includes OSGEP, TP53RK, TPRKB, and LAGE3. This complex catalyzes the threonylcarbamoyl adenosine (t6A) modification at position 37 of ANN-decoding tRNAs, a critical determinant of codon?Canticodon pairing accuracy. GON7 is essential for complex assembly and enzymatic activity; its loss disrupts tRNA modification, leading to ribosomal stalling, reduced translation fidelity, and activation of cellular stress responses. The KEOPS pathway is connected to mTORC1 signaling, which regulates translation initiation and elongation in response to nutrient status. Consequently, GON7 dysfunction contributes to pathologies such as Galloway-Mowat syndrome, primary microcephaly, and nephrotic syndrome, highlighting its role in development and kidney function.
Within the HEK293T background, GON7 knockout cells offer a tractable model to examine the consequences of impaired t6A modification on global protein synthesis and stress granule dynamics. The epithelial origin of HEK293T cells provides additional relevance for renal disease modeling. Researchers can apply western blotting to assess changes in KEOPS subunit stability, RT-qPCR to monitor tRNA expression patterns, and mass spectrometry to quantify t6A levels. Polysome profiling and stress granule analysis can further delineate translational defects, while immunofluorescence studies enable visualization of nucleolar alterations associated with KEOPS dysfunction.
This polyclonal knockout cell population is optimally suited for investigating the molecular underpinnings of translation regulation, tRNA modification biology, and KEOPS complex architecture. It enables functional complementation assays by reintroducing wild-type or mutant GON7 to dissect structure-function relationships. The model supports drug discovery efforts targeting translational control pathways and serves as a platform for studying related disorders, including Galloway-Mowat syndrome. For detailed technical specifications, pricing, and ordering information, please contact Ascent Research.
