The GON7 Knockout HeLa Polyclonal Cells product comprises a population of CRISPR/Cas9-edited HeLa cells with targeted disruption of the GON7 gene. This polyclonal knockout cell pool provides a genetically heterogeneous loss-of-function model for studying GON7-dependent processes. The CRISPR/Cas9-mediated gene disruption is achieved without isolation of single-cell clones, preserving population-level diversity while ablating GON7 protein expression. These polyclonal knockout cells are suitable for functional assays where uniform genetic background is not required, and the heterogeneous knockout enables assessment of gene essentiality in bulk populations.
The parental HeLa cell line is an extensively characterized human cervical epithelial adenocarcinoma model originally derived from a patient with HPV-18-associated cervical cancer. HeLa cells harbor integrated human papillomavirus type 18 sequences and exhibit dysregulated p53 and retinoblastoma protein pathways, contributing to their robust proliferation and widespread use in cancer biology. Their epithelial origin and transformed phenotype make them particularly relevant for investigating oncogenic signaling and tumor cell dependencies, providing a tractable system for gene knockout studies.
GON7 encodes a vital subunit of the KEOPS complex, which catalyzes the N6-threonylcarbamoyladenosine (t6A) modification of ANN-decoding tRNAs, including tRNA-Lys(UUU), tRNA-Thr(UGU), and tRNA-Asn(GUU). GON7 forms a macromolecular assembly with OSGEP, TP53RK, TPRKB, LAGE3, and YRDC, and its activity is modulated by upstream nutrient-sensing signals such as TORC1 kinase and amino acid availability. Loss of GON7 disrupts t6A modification, leading to impaired translation elongation and ribosomal fidelity, with downstream consequences on eEF1A-dependent peptide chain elongation and global protein synthesis. This positions GON7 as a critical node connecting nutrient signaling to translational control.
In HeLa cells, GON7 knockout generates a physiologically relevant model of defective tRNA modification within a cancer context. The HPV-18-driven transformation background permits investigation of how translational dysregulation intersects with oncogenic processes. Given GON7??s role in cellular growth regulation and its association with mTORC1 signaling, this knockout model is valuable for dissecting growth-related translational control mechanisms. Additionally, biallelic GON7 mutations cause Galloway-Mowat syndrome, a neurodevelopmental disorder, highlighting the model??s utility in studying the molecular basis of tRNA-modification-linked diseases beyond cancer.
Researchers can employ these polyclonal knockout cells in diverse assays, including Western blotting to confirm GON7 loss, HPLC or mass spectrometry-based tRNA modification profiling, puromycin incorporation assays to measure translation rates, and proliferation or cell cycle analyses by flow cytometry. Global translational changes can be assessed by RNA sequencing-based translational profiling. The heterogeneous knockout population is particularly suited for CRISPR-based genetic screens and pooled functional genomics studies. For detailed technical specifications, pricing, and ordering information, please contact Ascent Research.
