The EIF4G3 Knockout Huh-7 Polyclonal Cells are a ready-to-use CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma cell line. This product features a heterogeneous population of cells carrying targeted disruption of the EIF4G3 gene, achieved via CRISPR/Cas9-mediated genome editing. The polyclonal format provides a genetically diverse knockout pool suitable for pooled functional assays without requiring single-cell cloning. As a polyclonal knockout product, it is not a monoclonal cell line and does not guarantee biallelic or complete gene knockout in every cell. The knockout model enables loss-of-function studies of EIF4G3 in a biologically relevant hepatic cancer background.
The host cell line, Huh-7, is an adherent epithelial cell line originally established from a well-differentiated hepatocellular carcinoma. This tumorigenic line is extensively characterized and widely employed as a model system for liver function, hepatocyte biology, and hepatitis virus infection studies, particularly hepatitis C virus (HCV) replication. Huh-7 cells maintain key hepatic features and are permissive to HCV infection, making them a cornerstone for investigating host?Cvirus interactions and liver cancer pathogenesis. Their robust growth in culture and well-documented genetic background facilitate reproducible experimental designs and translational research.
EIF4G3 encodes a large scaffold protein central to the eIF4F translation initiation complex. It bridges eIF4E and eIF4A, and interacts with eIF3 and PABPC1 to recruit the 40S ribosomal subunit, activating cap-dependent translation of mRNAs encoding oncogenic factors like MYC and CCND1. Upstream, EIF4G3 is regulated by mTORC1 and PI3K-Akt signaling, as well as MAPK/ERK pathway inputs, and is cleaved by HCV NS3/4A protease to facilitate viral IRES-mediated translation. Additional interacting factors include MNK1 and the paralog eIF4G2 (DAP5), linking EIF4G3 to stress-responsive translation reprogramming.
In the Huh-7 hepatocellular carcinoma context, disruption of EIF4G3 has profound consequences. Loss of this scaffold protein dismantles the eIF4F complex, leading to reduced cap-dependent translation initiation, particularly of growth-promoting and survival mRNAs. This impairs cell proliferation, attenuates colony-forming ability, and may sensitize cells to apoptosis induced by therapeutic agents such as sorafenib. Given the reliance of HCC cells on elevated protein synthesis for sustained growth, EIF4G3 knockout creates a vulnerability that can be exploited to study translational dependency in liver cancer. Moreover, because HCV hijacks EIF4G3 for its own replication cycle, the knockout cells provide a clean background to dissect host factors essential for viral propagation, thereby aiding antiviral target identification.
This polyclonal knockout population enables investigation of translation control in liver cancer, screening for eIF4F complex inhibitors, and dissection of HCV-host interactions. Key assays include Western blotting, polysome profiling, cap-binding assays, RT-qPCR for MYC and CCND1, proliferation and colony formation assays, apoptosis analysis, and sorafenib sensitivity testing. HCV replication studies can also be performed. For further information or technical support, please contact Ascent Research.
