The E2F4 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma cell line. This product features targeted disruption of the E2F4 gene, resulting in a loss-of-function model for investigating the transcriptional repressor functions of E2F4.
Huh-7 is a well-differentiated hepatocellular carcinoma cell line originally isolated from a liver tumor of a Japanese male. This epithelial cell line is widely used in hepatocellular carcinoma research, drug metabolism studies, and hepatitis C virus replication assays. Huh-7 cells retain many hepatocyte-specific functions and are tumorigenic, making them a relevant model for studying liver cancer biology.
E2F4 is a member of the E2F transcription factor family and primarily functions as a transcriptional repressor of cell cycle genes. It frequently forms complexes with pocket proteins such as RB1, p107 (RBL1), and p130 (RBL2), and with co-repressors like HDAC1 and SIN3A, to inhibit the expression of targets including Cyclin A2 (CCNA2), Cyclin E1 (CCNE1), CDC6, and MCM genes. E2F4 activity is regulated upstream by Cyclin D/CDK4, TGF-beta, and Wnt/beta-catenin signaling, and it interacts with DP1 (TFDP1) and DP2 (TFDP2) to bind E2F sites in target promoters. As a component of the DREAM complex, E2F4 contributes to the maintenance of quiescence by repressing cell cycle progression.
In the context of Huh-7 hepatocellular carcinoma cells, which already exhibit dysregulated cell cycle control, knockout of E2F4 may relieve repression of proliferative genes, potentially enhancing cell cycle progression and providing a model to study the shift from quiescence to proliferation. This polyclonal knockout population enables investigation of E2F4’s tumor-suppressive functions and its interplay with the RB pathway in liver cancer. The mixed genotype of the polyclonal population mirrors the heterogeneity observed in tumors, offering a physiologically relevant system, although it does not guarantee complete gene inactivation in every cell.
This knockout model is suitable for a wide range of assays, including Western blotting and RT-qPCR to confirm target gene disruption and assess downstream effectors, cell proliferation assays (MTT, BrdU), flow cytometry for cell cycle analysis, and apoptosis detection. It also supports co-immunoprecipitation studies to probe E2F4-containing complexes and RNA-sequencing for transcriptome-wide insights into E2F4-dependent gene regulation. Applications extend to drug screening for hepatocellular carcinoma, functional analysis of the Hippo and TGF-beta pathways, and exploration of E2F4’s role in differentiation and tumor suppression. For further details and technical support, please contact Ascent Research.
