The GRPEL2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HT29 colorectal adenocarcinoma cells. They carry a targeted disruption of GRPEL2, which encodes a mitochondrial nucleotide exchange factor for mtHsp70, providing a loss-of-function model that avoids clonal bias.
The HT29 host cell line is an epithelial model originally from a 44-year-old female colorectal adenocarcinoma patient. It is extensively used for studying intestinal absorption, drug transport, and cancer biology, offering a physiologically relevant context for investigating mitochondrial function in colorectal tumor cells.
GRPEL2 functions as a nucleotide exchange factor for HSPA9 (mtHsp70), facilitating ADP release to drive the chaperone cycle essential for mitochondrial protein import, folding, and quality control. It interacts with HSPA9, DNAJA3, and the import complex, and is regulated by HSF1 and NRF2. Knockout disrupts client protein processing, lowers ATP, raises ROS, and activates UPRmt, with downstream effects on LONP1, CLPP, PINK1, and Parkin, linking to mitophagy.
In the HT29 background, GRPEL2 knockout reveals how mitochondrial proteostasis affects colorectal cancer cell fitness. These cells rely on mitochondrial metabolism for proliferation and stress adaptation; GRPEL2 loss sensitizes them to oxidative damage and metabolic crisis, making this model valuable for studying mitochondrial contributions to tumor progression.
Research applications include mitochondrial import studies, protein quality control, and UPRmt analysis. Compatible assays: Western blotting, ATP measurement, mitochondrial membrane potential (JC-1/TMRM), ROS detection (H2DCFDA), viability/apoptosis (MTT, Annexin V), Seahorse analysis, and co-immunoprecipitation. Applications span colorectal cancer, neurodegeneration, and drug screening for mitochondrial dysfunction. Contact Ascent Research for further details.
