The ATAD3A Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of human osteosarcoma 143B cells with targeted disruption of the ATAD3A gene. Unlike clonal cell lines, this polyclonal knockout model preserves phenotypic heterogeneity and reduces selection bias, providing a robust system for studying ATAD3A loss-of-function effects. This format enables consistent experimental outcomes across biological replicates and is well-suited for high-throughput screening applications.
The 143B cell line originated from a human osteosarcoma and has become a standard model in mitochondrial research due to its ample mitochondrial content and well-defined nuclear and mtDNA genomes. These adherent cells grow rapidly and are amenable to genetic manipulation, making them ideal for generating knockout models. Their tumorigenic origin also permits direct examination of mitochondrial roles in cancer cell proliferation and apoptosis.
ATAD3A is a mitochondrial inner membrane ATPase that regulates mitochondrial dynamics, cholesterol transport, and apoptosis. Upstream, MYC and Wnt/??-catenin signaling transcriptionally activate its expression, positioning it downstream of major oncogenic pathways. The protein interacts with VDAC1, HSP60, and Sam50 at contact sites, and with nucleoid proteins to influence mtDNA replication. Through BAX and BAK, ATAD3A modulates apoptotic pore formation. ATAD3A serves as a critical scaffold at mitochondrial contact sites, where it coordinates lipid transfer and interfacing with the endoplasmic reticulum. Its loss disrupts cholesterol trafficking, leading to metabolic stress and enhanced apoptosis.
In 143B osteosarcoma cells, ATAD3A knockout impairs mitochondrial integrity and energy production, sensitizing cells to apoptotic stimuli. Consequently, the cells exhibit reduced ATP levels and increased cytochrome c release upon stress. This model is valuable for dissecting how mitochondrial dysfunction affects cancer cell fitness, Wnt-driven mitochondrial regulation, and cholesterol-dependent survival signals. It also aids in exploring mechanisms relevant to ATAD3A-linked diseases such as Harel-Yoon syndrome. Thus, the cells serve as a platform to evaluate how ATAD3A interfaces with Wnt-dependent metabolism and apoptotic machinery in a bone cancer context.
Applications include western blotting for ATAD3A and apoptotic regulators (BAX, BAK), RT-qPCR for mitochondrial transcripts, ATP production and cholesterol quantification assays, cell viability testing, and mitochondrial membrane potential analysis using JC-1. The polyclonal knockout cells are suitable for drug sensitivity screening to identify compounds that target mitochondrial vulnerabilities in osteosarcoma. Furthermore, the model supports investigations into mitochondrial DNA replication defects and cholesterol-trafficking anomalies. Western blotting confirms ATAD3A depletion. For more details, contact Ascent Research.
