The Zbed6 Knockout AML12 Cell Line is a CRISPR/Cas9-edited knockout cell line designed to disrupt the Zbed6 gene, generating a loss-of-function model for investigating zinc finger BED domain-containing protein 6. This cell line provides a defined genetic background for studying Zbed6-mediated transcriptional repression and its downstream effects on insulin-like growth factor 2 (Igf2) expression. By abolishing Zbed6 function, the model enables researchers to examine de-repression of Igf2 and subsequent activation of IGF signaling cascades in a hepatocyte context. The knockout is achieved through CRISPR/Cas9-mediated gene disruption, ensuring stable and heritable ablation of the target locus without introducing exogenous sequences, making it suitable for a wide range of functional genomics and signaling studies.
The host AML12 cell line is a non-transformed mouse hepatocyte line derived from a transgenic mouse overexpressing human transforming growth factor alpha (TGF-??). AML12 cells retain characteristics of liver parenchymal cells, including expression of hepatocyte markers and metabolic activities, and are widely used to model hepatic metabolism, detoxification, and growth regulation. Their adherent growth and maintained differentiated functions make them an ideal platform for investigating liver-specific gene functions and signaling pathways. The TGF-?? transgene provides a proliferative signal that sustains the cells in culture without full transformation, preserving key hepatocyte features relevant to physiological and pathological studies.
Zbed6 functions as a transcriptional repressor that binds to a specific regulatory element within the Igf2 gene, negatively controlling its expression. Disruption of Zbed6 leads to elevated Igf2 transcription, which in turn engages the IGF1 receptor (Igf1r) and activates downstream signaling through Akt and mTOR pathways. This signaling axis is central to promoting cell growth, proliferation, and anabolic metabolism. Although upstream regulators of Zbed6 remain poorly characterized, it is likely to interact with transcriptional co-regulators to modulate chromatin structure and gene transcription. The Zbed6?CIgf2 axis is a key node in the IGF signaling network, with implications for muscle development and body composition, while in hepatocytes it may influence liver regeneration, metabolic homeostasis, and disease processes.
In the hepatocyte context, Zbed6 knockout enables dissection of Igf2-mediated growth regulation within the liver, where IGF signaling plays critical roles in development, regeneration, and metabolic control. The AML12 model allows investigation of how derepressed Igf2 affects hepatocyte proliferation, metabolism, and stress responses, offering insights into growth disorders and metabolic diseases. This cell line serves as a valuable tool for exploring the cell-autonomous effects of enhanced IGF signaling in liver parenchymal cells, potentially revealing mechanisms underlying hepatic hypertrophy or altered detoxification pathways. The intersection of Zbed6 function with hepatic physiology makes this model particularly relevant for studying liver-centric aspects of growth regulation and metabolic syndromes.
Researchers can employ the Zbed6 Knockout AML12 Cell Line in diverse experimental contexts, including quantitative analysis of Igf2 expression via qPCR, assessment of downstream signaling by Western blot for phosphorylated Akt and mTOR, and cell proliferation assays to quantify growth effects. Flow cytometry facilitates cell cycle profiling, while RNA-seq enables transcriptome-wide identification of Zbed6-dependent gene networks. These applications support investigations into IGF signaling, liver metabolism, muscle development, and growth disorders. The model is especially suited for comparative studies with wild-type AML12 cells to pinpoint Zbed6-specific contributions. For further information or to inquire about this product, please contact Ascent Research.
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