The E2F4 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-mediated gene disruption of the E2F4 transcriptional repressor in a polyclonal HT29 cell population. This polyclonal knockout cell pool offers a heterogeneous loss-of-function model, enabling study of E2F4-dependent regulatory networks without the bias of clonal selection.
HT29 is a human colorectal adenocarcinoma epithelial cell line derived from a primary tumor of a 44-year-old Caucasian female. It harbors mutant p53 and is widely used as a model for colorectal cancer studies. The cells exhibit epithelial morphology and are suitable for investigating tumor biology, signal transduction, and therapeutic responses.
E2F4 functions as a transcriptional repressor, forming complexes with pocket proteins (RB1, RBL1/p107, RBL2/p130) and DP cofactors (TFDP1, TFDP2) to silence E2F-responsive genes such as CCNE1, CDC6, and PCNA. This repressor activity is regulated by cyclin-dependent kinase-mediated phosphorylation and upstream signaling pathways including TGF-beta, which promotes E2F4-mediated G0/G1 arrest. The E2F4-DP-p130 complex binds to E2F target gene promoters, enforcing cell cycle exit. Knockout of E2F4 disrupts this repressive complex, potentially derepressing S-phase genes and altering cell cycle control.
In HT29 colorectal cancer cells, which already possess mutant p53 and dysregulated growth control, E2F4 knockout provides a powerful tool to dissect the contribution of transcriptional repression to tumor phenotypes. The loss of E2F4-mediated repression may exacerbate uncontrolled proliferation, migration, and invasion, mirroring aspects of aggressive colorectal cancer. This model allows researchers to examine how E2F4 loss interacts with existing oncogenic lesions, offering insights into the molecular evolution of colorectal malignancies and potential vulnerabilities.
This polyclonal knockout cell population is suitable for a broad range of functional studies, including cell cycle progression analysis by flow cytometry, proliferation assays (MTT/BrdU), colony formation, and drug sensitivity screens. Gene expression changes can be profiled via RNA-seq and RT-qPCR, while chromatin occupancy assessed by ChIP-qPCR. Migration and invasion assays can evaluate metastatic potential. The polyclonal nature provides a realistic model of heterogeneous tumor populations. For detailed experimental protocols or custom inquiries, please contact Ascent Research.
