The HERC2 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population in which the HERC2 gene is disrupted in the HT29 human colorectal adenocarcinoma cell line. This polyclonal format yields a heterogeneous pool of edited cells that collectively confer functional HERC2 loss, avoiding clonal artifacts and preserving cellular heterogeneity. The CRISPR/Cas9-mediated gene disruption provides a stable knockout model suitable for studying gene function in a physiologically relevant cancer cell context.
The HT29 cell line, derived from a primary colorectal adenocarcinoma of a 44-year-old female, displays adherent epithelial morphology and is extensively used as an intestinal epithelial model. HT29 cells maintain key features of colonic epithelium and are employed in colorectal cancer research to investigate tumor biology, drug responses, and signaling pathways. This host background enables direct analysis of HERC2 function in a malignancy-relevant cellular environment.
HERC2 encodes an E3 ubiquitin ligase that catalyzes ubiquitin transfer to substrate proteins, regulating their stability and function. It is integral to the DNA damage response, ubiquitinating repair factors including XPA, RPA, and the checkpoint mediator Claspin. HERC2 interacts with MDM2 to modulate p53 signaling, linking ubiquitin-mediated proteolysis to cell cycle control and apoptosis. Its activity is triggered by DNA damage-induced ATM/ATR kinase signaling and cellular stress, with downstream effects on targets such as OCA2. The ligase collaborates with UBE2D2 and forms complexes with multiple DNA repair proteins, positioning it as a critical coordinator of genome maintenance.
Disrupting HERC2 in HT29 cells creates a powerful model for dissecting its role in colorectal cancer. Given HERC2’s functions in DNA repair and p53 regulation, its loss may sensitize cells to genotoxic agents or impair cell cycle checkpoints, revealing cancer-specific vulnerabilities. Additionally, because HERC2 mutations are linked to Angelman syndrome and autism spectrum disorders, this model can help unravel shared molecular pathways between neurodevelopmental conditions and oncogenesis, despite the non-neuronal host.
Researchers can employ this polyclonal knockout model to investigate DNA damage response mechanisms, study ubiquitin-proteasome dynamics in colorectal cancer, and screen for modulators of HERC2 ligase activity. Compatible assays include Western blot and RT-qPCR for target gene analysis, co-immunoprecipitation to detect ubiquitination, flow cytometry for cell cycle profiling, DNA damage assays (comet assay, ??H2AX foci), and apoptosis or proliferation measurements. RNA-seq can further define transcriptomic changes. For technical inquiries, please contact Ascent Research.
