The RNF150 Knockout Huh-7 Cell Line is a CRISPR/Cas9-edited knockout cell line engineered to disrupt the RNF150 gene in the human Huh-7 hepatocellular carcinoma cell background. This product provides a loss-of-function model that enables precise investigation of RNF150-mediated ubiquitination processes and their functional consequences in a liver cancer context. The knockout cell line has been generated using CRISPR/Cas9 technology to introduce targeted gene disruption, resulting in ablation of full-length functional RNF150 protein expression, and offers a genetically defined system for studying the roles of this E3 ubiquitin ligase.
The host cell line, Huh-7, is a well-characterized human hepatocellular carcinoma cell line originally established from a liver tumor of a 57-year-old Japanese male. Huh-7 cells are widely employed as a model for hepatocyte biology and liver cancer research, as they retain many characteristics of differentiated hepatocytes while exhibiting aggressive tumorigenic properties. They harbor mutations in genes such as TP53 and ??-catenin and are responsive to a variety of stimuli, making them a versatile platform for dissecting oncogenic signaling and metabolic pathways. Their epithelial morphology, ease of culture, and susceptibility to gene editing further enhance their utility for functional genomic studies.
RNF150 encodes a RING finger E3 ubiquitin ligase that functions within the ubiquitin-proteasome system. The enzyme interacts with E2 ubiquitin-conjugating enzymes, ubiquitin, the proteasome, and substrate adaptor proteins to catalyze the transfer of ubiquitin moieties onto specific substrate proteins. This post-translational modification typically targets substrates for proteasomal degradation but can also modulate protein interactions, localization, or signaling activities. Although the upstream regulatory mechanisms and downstream substrate repertoire remain largely uncharacterized, RNF150 is believed to be controlled by transcriptional regulation and post-translational modifications. The pathway components involved in this system include E1 ubiquitin-activating enzyme, E2 conjugating enzymes, the 26S proteasome, and deubiquitinases that counterbalance ubiquitination. Knockout of RNF150 is expected to perturb the ubiquitination landscape, potentially stabilizing key regulatory proteins and altering downstream signaling cascades.
In the context of hepatocellular carcinoma, dysregulation of the ubiquitin-proteasome system is frequently observed and contributes to tumor initiation, progression, and drug resistance. The RNF150 Knockout Huh-7 Cell Line thus represents a valuable tool for elucidating the specific contribution of this E3 ligase to liver cancer biology. By comparing RNF150-deficient and parental Huh-7 cells, researchers can assess changes in cell proliferation, apoptosis, migration, and response to therapeutic agents. The model may reveal whether RNF150 acts as a tumor suppressor or oncogene in the liver, depending on the nature of its substrates, and could help identify novel biomarkers or therapeutic targets in hepatocellular carcinoma.
This knockout cell line supports a broad range of research applications, including functional characterization of RNF150 in hepatocellular carcinoma, identification of its ubiquitination substrates via co-immunoprecipitation and mass spectrometry-based proteomics, and investigation of protein degradation pathways. It is suitable for mechanistic studies linking RNF150 to signal transduction events, drug target validation through drug sensitivity testing, and phenotypic assays such as proliferation, apoptosis, and colony formation assays. Standard techniques like western blotting and RT?qPCR can be readily applied to confirm knockout efficiency and analyze downstream effects. For further details, please contact Ascent Research.
