This product is a polyclonal knockout cell population generated by CRISPR/Cas9-mediated gene disruption of EIF4G3 in K-562 cells. These polyclonal cells provide a loss-of-function model for studying EIF4G3-dependent translation initiation, enabling functional dissection of cap-dependent translation control without clonal variation.
K-562 is a human chronic myelogenous leukemia (CML) blast crisis cell line derived from a 53-year-old female. It harbors the Philadelphia chromosome (BCR-ABL1) and exhibits erythroleukemic features with pluripotent differentiation capacity into erythroid, granulocytic, and monocytic lineages, making it a well-established model for myeloid leukemia and hematopoietic differentiation research.
EIF4G3 is a scaffold protein of the eIF4F translation initiation complex, bridging the mRNA 5′ cap (via eIF4E) to the 40S ribosomal subunit (via eIF3) to drive cap-dependent translation. It also interacts with the RNA helicase eIF4A and the poly(A)-binding protein PABP, facilitating mRNA circularization and efficient ribosome recruitment. Its activity is controlled by mTORC1, which phosphorylates 4E-BP1 to liberate eIF4E, and by MAPK/ERK-MNK1 signaling, which phosphorylates eIF4E to enhance eIF4F assembly. Upstream growth factors such as insulin and EGF activate PI3K-AKT, converging on mTORC1 to stimulate EIF4G3-dependent translation. This scaffold selectively enhances translation of TOP mRNAs encoding proliferation regulators CCND1 and MYC, as well as anti-apoptotic proteins BCL2 and MCL1.
In K-562 cells, the BCR-ABL1 oncoprotein drives constitutive activation of mTOR and MAPK pathways, leading to elevated eIF4F complex assembly and enhanced translation of oncogenic mRNAs. Disrupting EIF4G3 in this polyclonal knockout population provides a powerful tool to investigate translation-dependent survival and proliferation mechanisms, enabling dissection of the contribution of cap-dependent translation to leukemic growth, differentiation, and drug resistance phenotypes.
This polyclonal knockout population is suitable for functional assays including Western blot analysis of EIF4G3, eIF4E, MYC, and CCND1; polysome profiling; puromycin incorporation assays to monitor global translation; proliferation (MTS/CCK-8) and apoptosis (Annexin V/PI flow cytometry) studies; and phospho-signaling analysis of phospho-4E-BP1 and phospho-S6K. It also supports drug sensitivity screening with mTOR inhibitors (e.g., rapamycin) or BCR-ABL inhibitors (imatinib). For further details, contact Ascent Research.
