The ATAD3A Knockout T-47D Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population in which the ATAD3A gene has been disrupted, creating a heterogeneous loss-of-function model system. This gene-edited cell pool is designed to circumvent clonal selection artifacts and provides a genetically diverse background for initial phenotypic screening, dose?Cresponse studies, and evaluation of target-gene function across a mixed allelic landscape.
The parental T-47D line was derived from a metastatic pleural effusion of an infiltrating ductal carcinoma of the breast and represents a well-characterized model of estrogen receptor-positive (ER+), progesterone receptor-positive (PR+) luminal A breast cancer. T-47D cells retain functional estrogen receptor alpha (ER??) signaling, exhibit hormone-dependent proliferation, and are extensively used to study endocrine therapy response, hormone-driven oncogenesis, and luminal subtype tumor biology.
ATAD3A encodes a mitochondrial inner membrane ATPase that nucleates the mitochondrial contact site and cristae organizing system (MICOS) through direct interaction with Mitofilin (IMMT), thereby maintaining crista junction architecture and inner membrane organization. ATAD3A also binds mitochondrial DNA (mtDNA) and supports nucleoid maintenance, and it partners with CYP11A1 to facilitate cholesterol transport for steroidogenesis. Expression of ATAD3A is transcriptionally activated by estrogen receptor alpha downstream of hormone stimulation and is further regulated by mTORC1 and PGC-1??. Loss of ATAD3A dysregulates mitochondrial dynamics factors OPA1, DRP1, and mitofusins MFN1/MFN2, leading to aberrant fusion/fission balance, and it sensitizes cells to intrinsic apoptosis through BAX/BAK activation. Consequently, ATAD3A disruption impairs cholesterol homeostasis, mtDNA integrity, and mitochondrial morphology.
In the context of T-47D ER+ breast cancer cells, ATAD3A knockout is expected to sever the link between hormone-driven proliferation and mitochondrial fitness. Because ATAD3A expression is estrogen-responsive and mTORC1-sensitive, its ablation may blunt metabolic adaptation to oncogenic signaling, increasing susceptibility to metabolic stress and impairing cellular energy production. Disrupted cholesterol handling may further compromise local steroidogenesis in this hormone-sensitive line, while mtDNA maintenance defects could provoke oxidative stress and inflammation. Thus, these polyclonal knockout cells offer a relevant model to dissect the mitochondrial underpinnings of luminal breast cancer and to identify synthetic lethal interactions or drug sensitivities that arise from ATAD3A loss.
Research applications include mitochondrial respirometry (Seahorse analysis) for bioenergetic profiling, mtDNA copy number quantification, and cholesterol measurement to assess metabolic perturbations. Apoptosis assays (Annexin V), fluorescence microscopy for mitochondrial morphology, and cell proliferation analyses enable functional interrogation of growth and death pathways. The heterogeneous knockout population is particularly suited for evaluating phenotypic heterogeneity, dose-dependent drug responses, and hormone signaling crosstalk, as well as for autophagy and ER stress pathway studies. For additional information and batch-specific quality control data, please contact Ascent Research.
