ID3 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout cell population derived from the human SK-HEP-1 cell line, featuring targeted disruption of the ID3 gene. This product provides a heterogeneous pool of cells with loss-of-function mutations in ID3, enabling functional studies without clonal isolation. The polyclonal format captures the diversity of editing outcomes, offering a robust model for investigating ID3-dependent processes in a liver adenocarcinoma background.
SK-HEP-1 is a human liver adenocarcinoma epithelial cell line originally established from the ascites of a patient with adenocarcinoma of the liver. It is widely employed as a hepatocellular carcinoma (HCC) model, facilitating research into liver cancer biology, tumor progression, and metastasis. The cells maintain epithelial morphology and are suitable for assays examining proliferation, migration, and signaling in the context of hepatic malignancy.
ID3 encodes inhibitor of DNA binding 3, a dominant-negative regulator of basic helix-loop-helix (bHLH) transcription factors. It functions by forming inactive heterodimers with E-proteins such as TCF3, TCF4, and TCF12, thereby preventing their DNA binding and attenuating target gene expression. ID3 is a key node in multiple signaling cascades, including TGF-?? and BMP pathways, where it is transcriptionally activated by SMAD2/SMAD3 complexes downstream of TGFB1 and BMP4. Additionally, ID3 is regulated by NOTCH1, E2F1, and TP53, and it directly modulates expression of downstream targets such as MYC, CCND1, CDKN1A, and BCL2, which govern cell cycle progression, apoptosis, and differentiation.
In SK-HEP-1 hepatocellular carcinoma cells, ID3 is known to promote proliferation and inhibit differentiation, partly through suppression of cyclin-dependent kinase inhibitor CDKN1A and upregulation of cyclin D1 (CCND1). Disruption of ID3 in this cell line thus provides a physiologically relevant loss-of-function model to dissect its role in HCC proliferation, survival, and metastatic potential. The polyclonal nature of the knockout allows assessment of ID3-dependent effects across a population, mimicking the heterogeneity often observed in tumor samples and enabling correlation of gene-dosage effects with phenotypic outcomes.
This ID3 knockout polyclonal cell population is suited for diverse experimental applications, including Western blotting, RT-qPCR, flow cytometric cell cycle analysis, migration and invasion assays, apoptosis assays, immunofluorescence, and TGF-??/BMP reporter gene assays. Researchers can utilize these cells to investigate TGF-??/BMP signaling in cancer, drug resistance mechanisms, metastasis, and the interplay between ID3 and other oncogenic pathways such as PI3K/Akt, Notch, and Wnt. The model is particularly valuable for co-culture studies, 3D spheroid assays, and high-content screening aimed at identifying compounds that modulate ID3 downstream networks. For additional technical details or ordering information, please contact Ascent Research.
