The ATAD3A Knockout SK-OV-3 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population engineered for targeted disruption of the human ATAD3A gene in the SK-OV-3 ovarian cancer background. This genetically modified cell pool provides a loss-of-function model suitable for investigating ATAD3A-dependent mitochondrial processes in a disease-relevant cellular context. The polyclonal knockout product format preserves the genetic heterogeneity of the edited cell population, enabling robust assessment of ATAD3A ablation across a mixed genotype landscape without selection for single-cell clones. This versatile tool is intended to facilitate mechanistic studies of mitochondrial biology, cholesterol homeostasis, and oncogenic signaling in epithelial ovarian adenocarcinoma.
The host SK-OV-3 cell line is derived from the ascites of a 64-year-old female patient with ovarian serous adenocarcinoma, a subtype characterized by aggressive peritoneal spread and frequent chemoresistance. This adherent cell model harbors a TP53 mutation and exhibits HER2 positivity, reflecting genomic alterations commonly observed in high-grade serous ovarian carcinomas. SK-OV-3 cells serve as a well-established platform for tumor cell biology, drug response profiling, and dissecting pathways that drive ovarian cancer progression. The integration of CRISPR/Cas9-mediated ATAD3A disruption into this genetically defined background allows direct interrogation of mitochondrial contributions to ovarian cancer pathophysiology.
ATAD3A encodes an inner mitochondrial membrane ATPase critically regulating mitochondrial dynamics, cholesterol transport, and mtDNA nucleoid organization. It interacts with ATAD3B, prohibitins, MICOS components Mic10/Mic60, and StAR to sustain mitochondrial architecture. Upstream signals from ESR1, AR, p53, MYC, and mTORC1 modulate ATAD3A activity, connecting it to oncogenic pathways. Knockout of ATAD3A perturbs downstream targets DRP1, FIS1, and cholesterol synthesis enzymes, reduces mtDNA copy number, and disrupts ER?Cmitochondria contacts, which can sensitize cells to BAX/BAK-mediated apoptosis. Thus, ATAD3A operates at the intersection of organelle communication and metabolic control.
In the TP53-mutant, HER2-positive SK-OV-3 background, ATAD3A knockout enables dissection of mitochondrial contributions to ovarian cancer proliferation, metabolic adaptation, and apoptotic resistance. This polyclonal model is well-suited for exploring p53-independent vulnerabilities and the interplay between oncogenic signaling and cholesterol trafficking. The heterogeneous knockout population mirrors tumor diversity, facilitating the identification of synthetic lethal interactions or resistance mechanisms dependent on ATAD3A.
Applications include western blotting, RT-qPCR, immunofluorescence, and flow cytometry for verifying ATAD3A ablation and monitoring mitochondrial morphology. Cholesterol quantification and mtDNA copy number assays assess metabolic and genomic consequences. Cell proliferation, apoptosis, and migration/invasion assays further characterize phenotypic outcomes. Co-immunoprecipitation can probe interactions with ATAD3B, StAR, and prohibitins. Pharmacological inhibitors of mTORC1 or AR may be used to interrogate regulatory nodes. For additional information, contact Ascent Research.
