The EEF1D Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population that disrupts the EEF1D gene in the HT29 human colorectal adenocarcinoma cell line. This product provides a loss-of-function model for investigating the role of the EEF1D-encoded protein in translational control and stress responses, without implying a specific allelic configuration or clonal homogeneity. The polyclonal format preserves population-level heterogeneity while ensuring target-gene disruption, making it suitable for pooled functional studies.
HT29 cells were originally isolated from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female and are widely employed as a model of intestinal epithelial biology. These cells retain the capacity to differentiate into mucus-secreting and absorptive phenotypes, recapitulating key features of the colonic epithelium. Their adherent growth, well-characterized signaling networks, and relevance to colorectal cancer research make them a robust host for gene-editing studies focused on tumor biology and therapeutic resistance.
EEF1D encodes the guanine nucleotide exchange factor (GEF) for eEF1A, loading eEF1A with GTP to enable aminoacyl-tRNA binding during translational elongation. Beyond its canonical role, EEF1D also mediates heat shock-induced HSP70 (HSPA1A) expression via HSF1 independently of GEF activity. EEF1D is regulated by CK2 phosphorylation and mTORC1 signaling, and it interacts with eEF1A, EEF1B??, EEF1B??, and valyl-tRNA synthetase. Through these complexes, it influences ribosomal protein S6 and global protein synthesis, linking nutrient and stress cues to translational output.
In HT29 colorectal adenocarcinoma cells, EEF1D knockout provides a model to examine how translational elongation contributes to cancer phenotypes such as uncontrolled proliferation and drug resistance. HT29 cells display mTOR pathway activation typical of colorectal tumors, making them well-suited to study EEF1D??s integration of growth and stress signals. Disruption of EEF1D may alter HSP70-mediated cytoprotection and ribosomal S6 phosphorylation, thereby affecting cell survival under therapeutic pressure.
The EEF1D Knockout HT29 Polyclonal Cells support diverse functional assays including polysome profiling and puromycin incorporation to quantify translation rates, heat shock treatments coupled with HSP70 immunodetection, and cell viability or colony formation assays to investigate drug sensitivity. Additional applications encompass TEER measurements for barrier function, flow cytometry-based analyses, and functional genomics screening. This model enables dissection of translational control in colorectal cancer and stress biology. For further information, please contact Ascent Research.
