The HECTD1 Knockout HeLa Polyclonal Cells are a polyclonal knockout cell population derived from the HeLa human cervical carcinoma cell line, genetically engineered using CRISPR/Cas9-mediated gene disruption to ablate HECTD1 expression. This loss-of-function model enables systematic investigation of the HECT domain-containing E3 ubiquitin ligase HECTD1 in a tumorigenic background.
The host HeLa cell line is an HPV18-positive cervical adenocarcinoma-derived immortalized cell line, widely employed in cancer research for its robust growth and well-characterized signaling networks. Its genetic stability and extensive historical use facilitate reproducible studies in cellular transformation, migration, and drug response.
HECTD1 encodes a HECT-domain E3 ubiquitin ligase that regulates protein turnover and signaling by targeting substrates such as HSP90AA1, PIAS4, ??-catenin, and Smad7 for ubiquitination and degradation. It is activated by cellular stress stimuli and transcriptionally regulated by heat shock factor 1 (HSF1) and MYC. Through ubiquitination of HSP90AA1, HECTD1 modulates chaperone activity and protein quality control within the HSP90 cycle. Additionally, HECTD1 interfaces with Wnt/??-catenin signaling by interacting with adenomatous polyposis coli (APC) and controlling ??-catenin stability, affecting TCF/LEF-dependent transcription. It also impacts TGF-?? signaling via Smad7 and participates in NF-??B modulation through I??B?? regulation, collectively influencing cell proliferation, migration, and stress adaptation.
In the HeLa cell context, disruption of HECTD1 provides a potent system to dissect ubiquitin-mediated regulation of oncogenic pathways. HeLa cells exhibit elevated HSP90 activity and aberrant Wnt signaling, making them ideal for studying HECTD1??s role in chaperone-dependent protein stabilization and ??-catenin-driven transcription. This model is particularly relevant for exploring mechanisms of cervical cancer progression, where protein quality control and signaling crosstalk are frequently dysregulated.
This knockout product supports diverse research applications including ubiquitination assays to identify novel substrates, co-immunoprecipitation (Co-IP) to map HECTD1 interaction networks, and Western blot analysis of ubiquitinated proteins. It is suitable for functional studies using HSP90 chaperone activity assays, ??-catenin reporter systems such as TOPFlash/FOPFlash, and Boyden chamber-based migration and invasion assays. The polyclonal population also facilitates drug sensitivity testing with HSP90 inhibitors (e.g., 17-AAG) or proteasome inhibitors (MG132), and quantitative RT-PCR for Wnt target gene expression. For further inquiries, please contact Ascent Research.
