The ADCK2 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population derived from the HCT 116 human colorectal carcinoma cell line, designed to disrupt the ADCK2 gene. This heterogeneous knockout population is ideal for pooled functional studies and bulk biochemical analyses without clonal selection artifacts.
The HCT 116 host cell line is a widely used colorectal adenocarcinoma epithelial model characterized by microsatellite instability due to a homozygous mutation in the MLH1 mismatch repair gene, wild-type TP53, and an oncogenic KRAS G13D mutation. These genetic alterations recapitulate key features of a subset of human colorectal cancers, including defective DNA repair and constitutive mitogenic signaling. The cell line??s robust growth properties and well-documented molecular landscape make it an ideal background for studying metabolic pathways and their impact on cancer cell behavior.
ADCK2 encodes a mitochondrial atypical kinase that acts as a regulatory hub in the coenzyme Q biosynthetic pathway. Transcriptionally controlled by NRF1, NRF2, and PGC-1??, and imported into mitochondria via specialized receptors, ADCK2 phosphorylates and stabilizes COQ3, COQ5, and COQ7, and interacts with COQ9. This stabilization promotes CoQ10 synthesis, facilitating electron transfer through the respiratory chain. ADCK2 loss disrupts this complex, reducing CoQ10, impairing electron transport, and altering ATP production.
In HCT 116 colorectal cancer cells, ADCK2-mediated CoQ10 biosynthesis is particularly relevant due to metabolic reprogramming that accompanies tumorigenesis. Colorectal tumors often upregulate mitochondrial biogenesis, and CoQ10 is critical for electron transport and antioxidant defense. ADCK2 knockout in this MSI-high, KRAS-mutant background reveals how mitochondrial CoQ10 contributes to cancer cell fitness and chemoresistance, and it provides a model for coenzyme Q10 deficiency disorders. This tool enables dissection of the intersection between mitochondrial metabolism and oncogenic signaling.
Researchers can employ these polyclonal knockout cells for Western blotting of COQ proteins, LC?MS-based CoQ10 quantification, Seahorse metabolic flux analysis, blue native PAGE for mitochondrial supercomplexes, ATP colorimetric assays, and flow cytometry-based apoptosis assays. This model supports pooled functional screens and detailed coenzyme Q pathway characterization. It enables mechanistic studies of mitochondrial kinase signaling and pharmacologic screening for modulators of CoQ10 metabolism. For further information, please contact Ascent Research.
