ARL6IP1 Knockout HT29 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of human colorectal adenocarcinoma HT29 cells with disrupted ARL6IP1 expression. ARL6IP1 encodes an endoplasmic reticulum (ER)-resident protein involved in ER morphology, protein trafficking, and anti-apoptotic signaling. Loss of ARL6IP1 sensitizes cells to ER stress-induced apoptosis, providing a valuable model for studying ER stress pathways and apoptotic regulation in a colorectal cancer background.
The HT29 cell line, derived from a colorectal adenocarcinoma of a 44-year-old female, is a widely used model for colorectal cancer research, including tumor progression, metastasis, and drug response. HT29 cells also differentiate into enterocyte-like monolayers, enabling intestinal barrier function studies. They carry relevant oncogenic mutations, making them a clinically relevant platform for gene function analysis.
ARL6IP1 localizes to the ER, where it interacts with BCL2 to inhibit apoptosis and modulate the unfolded protein response (UPR). Under ER stress, ARL6IP1 reduces activation of PERK and IRE1, limiting induction of pro-apoptotic factors CHOP and GRP78. Its knockout releases BAX-mediated mitochondrial permeabilization, triggering Caspase-3 and Caspase-9 activation. ARL6IP1 also associates with ER morphology regulators ATL1 and RTN4, and is regulated by the small GTPase ARL6. Crosstalk with autophagy is evidenced by changes in LC3 levels upon stress.
In HT29 cells, ARL6IP1 knockout creates a model to examine how ER stress influences colorectal cancer cell survival. Given that colorectal tumors face hypoxia and nutrient deprivation, UPR modulation is a potential therapeutic target. This knockout enables assessment of proliferation, clonogenic growth, and chemosensitivity to agents like 5-fluorouracil under ER stress. It also aids in studying the role of ARL6IP1 in intestinal epithelial barrier integrity during inflammatory or stress conditions.
Researchers can apply this model for studying ER stress-mediated apoptosis using Western blotting, cell viability assays under tunicamycin or thapsigargin treatment, and caspase activity assays. Transcriptional analysis of CHOP and GRP78 by qPCR monitors UPR activation, while co-immunoprecipitation validates ARL6IP1-BCL2 interaction. Immunofluorescence visualizes ER morphology changes, and flow cytometry quantifies apoptosis via annexin V staining. This model also supports drug screening for ER stress modulators in colorectal cancer. For further information, please contact Ascent Research.
