The BTBD10 Knockout HeLa Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the BTBD10 gene in the HeLa human cervical adenocarcinoma cell line. This genetically modified pool serves as a versatile loss-of-function model for investigating the biological roles of the BTB domain-containing protein 10 in cellular ubiquitination processes. The polyclonal nature of the knockout population captures a range of editing events, offering a robust system for functional studies without the clonal variability associated with single-cell-derived lines. Researchers can employ this tool to dissect the contributions of BTBD10 to substrate-specific degradation within the ubiquitin-proteasome system.
HeLa cells, the host background, are an immortalized epithelial cell line originally derived from a cervical adenocarcinoma. These cells are widely utilized in cancer research due to their robust growth, ease of manipulation, and well-characterized signaling networks. The cervical origin makes this line particularly relevant for studying tumorigenic mechanisms and testing therapeutic interventions targeting cervical and other solid tumors. By introducing BTBD10 knockout into this established background, the model enables direct examination of gene function in a disease-relevant context.
BTBD10 encodes a substrate recognition adaptor of CUL3-based E3 ubiquitin ligase complexes. The protein assembles with the scaffold CUL3 and the RING finger protein RBX1 to form an active ubiquitin ligase that transfers ubiquitin from E2 conjugating enzymes to specific substrates, tagging them for 26S proteasome-mediated degradation. Through this function, BTBD10 regulates key cellular processes including cell cycle progression and apoptosis. The activity of this complex is modulated by CUL3 expression levels, neddylation status of CUL3, and transcription factors of the BTB family. Interacting partners include E2 enzymes, proteasome subunits, and alternative BTB adaptors, placing BTBD10 at a critical node of the ubiquitin-proteasome pathway.
In HeLa cervical cancer cells, BTBD10-dependent ubiquitination likely controls the turnover of proteins that govern proliferation and survival. Disruption of BTBD10 expression is expected to stabilize its downstream substrates, potentially revealing novel regulators of oncogenic signaling. This model provides a direct means to assess the functional consequences of impaired CUL3-BTB complex activity in a cervical carcinoma background, facilitating the identification of substrates whose accumulation or depletion alters cancer cell phenotypes. The knockout cells are thus a powerful resource for exploring how ubiquitin ligase adaptors contribute to tumor cell biology.
Applications of the BTBD10 Knockout HeLa Polyclonal Cells span mechanistic studies of ubiquitin-dependent proteolysis, substrate profiling via ubiquitination assays and western blotting, and investigation of protein?Cprotein interactions through co-immunoprecipitation of the CUL3?CBTBD10 complex. The model is suitable for phenotypic assays such as MTT-based cell viability, Annexin V apoptosis detection, and flow cytometric cell cycle analysis, alongside proteasome activity measurements and RT-qPCR evaluation of gene expression changes. These applications enable screening for modulators of the ubiquitin-proteasome system and functional genomics initiatives aimed at characterizing BTBD10 targets in cervical cancer. For further inquiries, please contact Ascent Research.
