The HERC4 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, designed to disrupt HERC4 gene function. This polyclonal knockout model comprises a heterogeneous pool of cells with targeted gene disruption, enabling population-level studies of loss-of-function effects in a cancer-relevant epithelial context. The polyclonal format maintains the inherent genetic variability of the host cell line while providing robust knockout representation for reproducible experimental analyses.
The HT29 cell line is an established model of human intestinal epithelium, originating from a colorectal adenocarcinoma and exhibiting adherent epithelial morphology. It harbors inactivating mutations in the tumor suppressor p53 and the adenomatous polyposis coli (APC) gene, two frequent genetic lesions in colorectal carcinogenesis. HT29 cells are capable of enterocytic differentiation and formation of polarized monolayers, making them a versatile platform for studying cancer cell biology, signal transduction, and therapeutic responses.
HERC4 is an E3 ubiquitin-protein ligase that targets substrates for proteasomal degradation. In JAK-STAT signaling, HERC4 ubiquitinates PIAS1, a SUMO E3 ligase and STAT1 inhibitor, promoting PIAS1 degradation by the proteasome. This releases STAT1-mediated transcriptional activity, boosting expression of interferon-responsive genes such as IRF1. HERC4 functions downstream of interferon-gamma receptor (IFNGR1) and the kinases JAK1 and JAK2, which activate STAT1. Thus, HERC4 modulates JAK-STAT signaling by controlling PIAS1 stability, linking ubiquitination to transcriptional output.
Knockout of HERC4 in the HT29 colorectal adenocarcinoma background provides a system to dissect how E3 ligase dysfunction impacts tumor cell behavior. With the HT29 line’s mutant APC and p53, HERC4 disruption can reveal its contribution to uncontrolled proliferation and altered cytokine responses. By removing HERC4, researchers can examine shifts in PIAS1 protein levels, STAT1 activation, and downstream gene expression, linking ubiquitin-proteasome activity to colorectal cancer phenotypes. The polyclonal knockout population enables the study of JAK-STAT pathway alterations in a heterogeneous setting, mirroring the genetic diversity of tumors and allowing identification of clonal variation in signaling sensitivity.
These HERC4 knockout polyclonal cells are suitable for a range of experimental applications, including PIAS1 stability assays via Western blotting, STAT1 luciferase reporter assays, and RT-qPCR analysis of IRF1 and other targets. Cell proliferation and interferon-gamma sensitivity assays can dissect the role of HERC4 in growth control and cytokine responsiveness. Additionally, the cells serve as a model for studying ubiquitin-proteasome pathway regulation in colorectal cancer and for exploring JAK-STAT signaling dynamics. For further details or assistance, please contact Ascent Research.
