GTPBP10 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population harboring targeted disruption of the GTPBP10 gene, a nucleolar GTPase essential for 60S ribosomal subunit maturation. This product is supplied as a polyclonal pool, providing a robust loss-of-function model while avoiding clonal selection artifacts that may confound functional genomic studies. The knockout cells are designed for investigations into ribosome biogenesis, nucleolar stress responses, and their implications in cancer and ribosomopathies, leveraging the well-characterized HEK293T background.
The parental HEK293T cell line is derived from human embryonic kidney cells and stably expresses the SV40 large T antigen, which facilitates high-level protein expression and efficient viral production. These features make HEK293T a versatile platform for studying nucleolar biology, as the cells exhibit rapid growth and active ribosome synthesis. The ease of transfection and transduction further enhances their utility for complementation assays and downstream pathway analysis in the context of GTPBP10 deficiency.
GTPBP10 encodes a nucleolar GTPase that promotes the maturation of the pre-60S ribosomal subunit by interacting with key assembly factors and ribosomal proteins. Mechanistically, GTPBP10 forms complexes with RPL5, RPL11, and NPM1 within the nucleolus, and its GTPase activity is required for proper rRNA processing and pre-60S export. The gene is transcriptionally activated by c-Myc, linking ribosome production to growth signals, and its disruption leads to impaired ribosomal protein synthesis. Knockout of GTPBP10 triggers nucleolar stress, which may activate p53-dependent pathways through sequestration of RPL5 and RPL11, thereby coupling ribosome biogenesis to cell cycle control.
In the HEK293T context, loss of GTPBP10 profoundly disrupts ribosome assembly, leading to accumulation of pre-rRNA intermediates and nucleolar morphological changes. This polyclonal knockout population serves as a physiologically relevant model to dissect the interplay between ribosome biogenesis, nucleolar integrity, and cellular proliferation. Notably, the absence of GTPBP10 sensitizes cells to impaired translation and may reveal synthetic vulnerabilities relevant to cancer therapy, particularly in tumors driven by hyperactive c-Myc signaling. The model enables detailed examination of how nucleolar stress is transmitted to downstream effectors, including p53 and ribosomal proteins.
Key applications include monitoring pre-rRNA processing and ribosomal protein levels via RT-qPCR and Western blotting, visualizing nucleolar disruption by immunofluorescence, and assessing translation efficiency through polysome profiling. Co-immunoprecipitation assays can be employed to study GTPBP10 interactions with pre-60S factors and ribosomal proteins, while GTPase activity measurements provide mechanistic insight. Additionally, the cells are ideal for p53 activation and viability assays to explore stress responses. For complete product information and support, please contact Ascent Research.
