ATAD3A Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line. This product provides a genetically perturbed model in which the ATAD3A gene has been disrupted via CRISPR/Cas9-mediated gene targeting, resulting in a heterogeneous pool of cells carrying diverse loss-of-function mutations. As a polyclonal population, it reflects the collective effects of ATAD3A ablation without clonal selection, making it suitable for bulk population-based assays and functional genomics studies. The cells are supplied as a ready-to-use knockout reagent for investigating ATAD3A-dependent processes in cancer biology.
The parental HCT 116 cell line is an epithelial colorectal carcinoma model established from a human male with microsatellite instability (MSI-high) and activating mutations in KRAS (G13D) and PIK3CA (H1047R). These genetic alterations drive constitutive proliferative and survival signaling, rendering HCT 116 cells a widely used system for colorectal cancer research, drug sensitivity profiling, and signal transduction studies. The defined genetic background provides a consistent platform to assess the functional consequences of ATAD3A disruption in a tumorigenic context with known oncogenic drivers.
ATAD3A encodes a mitochondrial AAA ATPase that localizes predominantly at endoplasmic reticulum?Cmitochondria contact sites, where it orchestrates nucleoid organization, cholesterol trafficking, and cristae architecture. ATAD3A interacts with inner mitochondrial membrane components SAM50 and MIC60 and associates with VDAC1 at outer membrane contact points, facilitating cholesterol import and regulating mitochondrial fission by modulating DRP1 activity. Under cholesterol depletion or PGC-1??-mediated biogenesis, ATAD3A function is augmented, while mitophagy inducers and ER stress can alter its expression or assembly. Downstream, ATAD3A tightly controls mtDNA maintenance via TFAM and governs apoptosis by sequestering cytochrome c; its loss triggers cytochrome c release, caspase-3 activation, and BNIP3-mediated mitophagy. Thus, ATAD3A sits at the nexus of metabolic and apoptotic signaling, coupling lipid homeostasis to mitochondrial ultrastructure and cell fate decisions.
In the HCT 116 background, ATAD3A disruption unmasks vulnerabilities arising from the intersection of altered cholesterol metabolism, mitochondrial dynamics, and apoptotic thresholds that are characteristic of MSI-high colorectal cancers. The coexistence of KRAS and PIK3CA mutations generates a metabolic and survival milieu in which ATAD3A-dependent contact site integrity may be critical for sustaining proliferation and evading cell death. Consequently, this knockout model enables dissection of mitochondrial contributions to oncogenic transformation, identification of synthetic lethal interactions with cholesterol synthesis inhibitors or mTOR pathway modulators, and exploration of mitochondrial priming for apoptosis-based therapies.
Researchers can employ these polyclonal knockout cells in a range of targeted assays. Western blotting and immunofluorescence validate loss of ATAD3A protein and assess downstream effectors such as DRP1 and cytochrome c localization. Co-immunoprecipitation studies examine disrupted interactions with SAM50, MIC60, and VDAC1. mtDNA copy number qPCR measures nucleoid maintenance defects, while cholesterol uptake assays quantify transport impairments. Apoptosis detection by Annexin V/PI staining and caspase-3 activity assays reveals sensitivity to intrinsic cell death, and Seahorse metabolic flux analysis evaluates mitochondrial respiratory capacity. These applications make the ATAD3A Knockout HCT 116 Polyclonal Cells a versatile tool for mechanistic oncology and mitochondrial biology research. For additional information or custom inquiries, please contact Ascent Research.
