The HERPUD2 Knockout HT29 Polyclonal Cells product consists of a heterogeneous CRISPR/Cas9-edited HT29 colorectal adenocarcinoma cell population with targeted loss of HERPUD2 function. This polyclonal knockout model enables investigation of endoplasmic reticulum stress and ER-associated degradation (ERAD) pathways in an epithelial context. The cryopreserved stock retains the parental line??s ability to undergo partial enterocytic differentiation upon confluence or treatment with sodium butyrate.
HT29 cells were originally isolated from a primary colorectal adenocarcinoma of a 44-year-old female and are extensively used as a colorectal cancer model. Under standard conditions, they grow as an undifferentiated epithelial monolayer, but can undergo enterocytic differentiation upon confluence or treatment with sodium butyrate. The parental line harbors mutations in APC and TP53, making it a valuable system for investigating oncogenic signaling, drug responses, and differentiation biology.
HERPUD2 encodes an ER-resident protein that functions as a key negative regulator of the unfolded protein response (UPR). Upon ER stress induction by agents like tunicamycin or thapsigargin, HERPUD2 is upregulated via ATF6 and XBP1. It then recruits the HRD1/SYVN1 ubiquitin ligase complex, together with VCP/p97, SEL1L, and DERL1, to promote degradation of misfolded proteins. This activity attenuates IRE1 and PERK signaling, lowers CHOP (DDIT3) expression, and suppresses pro-apoptotic mediators such as Bim and caspases, thereby protecting cells from ER stress-induced cell death.
The HT29 colorectal cancer background provides a relevant context for studying HERPUD2, as tumor cells frequently encounter proteotoxic stress and rely on UPR signaling for survival. HERPUD2 disruption in this polyclonal pool enhances sensitivity to ER stress inducers and proteasome inhibitors, highlighting its prosurvival function. Additionally, the partial differentiation capacity of HT29 cells allows exploration of HERPUD2??s involvement in enterocytic differentiation under stress conditions.
This polyclonal knockout product is optimized for functional assays such as Western blotting and RT-qPCR to monitor UPR markers (BiP/GRP78, CHOP), cell viability and apoptosis assays under ER stress challenge (MTT, Annexin V, caspase-3/7), co-immunoprecipitation of HERPUD2 with HRD1, and cycloheximide chase analyses of protein turnover. Applications include drug sensitivity screening, mechanistic dissection of ER stress signaling in colorectal cancer, and crosstalk studies between UPR and differentiation. For further details or custom requests, please contact Ascent Research.
