The IRGQ Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma epithelial cell line, engineered for targeted disruption of the IRGQ gene. This polyclonal population provides a genetically heterogeneous loss-of-function model enabling interrogation of mitophagy and tumor suppressor pathways. Use of CRISPR/Cas9-mediated gene disruption ensures ablation of IRGQ protein function for studying impaired mitochondrial quality control.
HT29 cells, isolated from a primary colorectal adenocarcinoma, are widely used in colorectal cancer and epithelial biology research. They display epithelial morphology and harbor mutant APC and TP53 (R273H) while retaining wild-type KRAS, making them a relevant model for studying intestinal epithelial differentiation, cancer progression, and drug absorption.
IRGQ acts as a mitophagy receptor that targets damaged mitochondria for autophagic degradation, functioning as a tumor suppressor in colorectal cancer. It directly binds ATG8 family members LC3B, GABARAP, GABARAPL1, and GABARAPL2 to recruit autophagosomes. IRGQ is regulated by cellular stress signals such as hypoxia, with potential involvement of HIF1A and TFEB, and operates within the autophagy pathway alongside the ULK1-ATG13-FIP200 complex, PI3KC3, and ATG5-ATG12-ATG16L1 conjugation systems. Loss of IRGQ disrupts mitophagy, leading to mitochondrial accumulation, elevated ROS, and enhanced cancer cell proliferation and migration.
In HT29 cells with APC and TP53 mutations, IRGQ knockout provides a tool to study how mitophagy deficiency cooperates with oncogenic pathways in colorectal cancer. This model facilitates investigation of mTOR signaling crosstalk and mitochondrial quality control in a genetic background representative of colorectal tumors.
Applications include western blotting for LC3-I/II and p62, immunofluorescence for LC3 puncta and mitochondrial markers, flow cytometry for mitochondrial mass and ROS, RT-qPCR, proliferation (MTT, colony formation) and migration (Transwell, wound healing) assays, co-immunoprecipitation for ATG8 interactions, Seahorse mitochondrial stress tests, and xenograft tumor growth. These cells are suitable for autophagy-targeted drug development and signaling studies. For technical support, contact Ascent Research.
