The EEF1A2 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma cell line, designed to disrupt the EEF1A2 gene encoding the alpha 2 isoform of eukaryotic elongation factor 1. This polyclonal population offers a heterogeneous pool of loss-of-function alleles, enabling robust functional interrogation of EEF1A2 in a liver cancer context without clonal selection bias.
The parental Huh-7 cell line originates from a well-differentiated hepatocellular carcinoma of a 57-year-old Japanese male and is widely utilized for studying hepatic metabolism, detoxification pathways, and protein synthesis. Huh-7 cells retain characteristic epithelial morphology and express liver-specific markers, making them a suitable model for investigating molecular mechanisms underlying hepatocarcinogenesis and liver cell biology.
EEF1A2 functions as a translation elongation factor that delivers aminoacyl-tRNAs to the ribosome, a process tightly regulated by upstream signals such as MYC, E2F1, mTORC1/S6K1, and MAPK/ERK. It physically interacts with aminoacyl-tRNA, ribosomes, actin, PI4KIII??, and G proteins, and its activity promotes global protein synthesis, actin cytoskeleton organization, and Akt activation, leading to the upregulation of anti-apoptotic Bcl-2 family proteins. Through these interactions, EEF1A2 integrates nutrient and growth factor signaling via the mTOR and PI3K-Akt pathways to control cell growth and survival.
In Huh-7 hepatocellular carcinoma cells, disruption of EEF1A2 is expected to impair translation elongation and perturb actin dynamics, potentially reducing cell proliferation and enhancing apoptosis susceptibility. Given the established anti-apoptotic function of EEF1A2 in cancer, this knockout model provides a valuable tool to dissect the dependency of liver cancer cells on sustained protein synthesis and cytoskeletal integrity, and to explore how loss of EEF1A2 affects signaling through Akt-dependent survival mechanisms.
Researchers can employ this knockout polyclonal population in diverse applications, including investigating the role of EEF1A2 in hepatocellular carcinoma progression, screening for small-molecule inhibitors targeting translation elongation in liver cancer, and exploring non-canonical functions of eEF1A2 beyond protein synthesis. Representative assays include western blotting and RT-qPCR for expression analysis, polysome profiling to assess global translation, MTT/CCK-8 and colony formation assays for proliferation, Annexin V staining for apoptosis, and phalloidin staining for actin cytoskeleton visualization. This product is also suitable for tumor xenograft studies to evaluate metastatic potential. For additional information or custom inquiries, please contact Ascent Research.
