The ATAD3A Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt ATAD3A expression in a human embryonic kidney background. This polyclonal pool contains a heterogeneous mix of edited alleles, providing a robust loss-of-function model while avoiding clonal selection biases. The product serves as a versatile tool for dissecting ATAD3A-dependent mitochondrial processes in a highly transfectable host.
Derived from human embryonic kidney cells, the HEK293T line is stably transformed with adenovirus 5 DNA and constitutively expresses the SV40 large T antigen, enabling episomal replication of plasmids with the SV40 origin. Renowned for its exceptional transfectability, HEK293T is a standard host for recombinant protein production, lentiviral packaging, and functional genomics. Its epithelial origin and rapid proliferation facilitate high-resolution imaging and biochemical analyses of mitochondrial parameters, making it ideal for studying organellar biology.
ATAD3A is a nuclear-encoded mitochondrial inner membrane protein critical for cristae organization, cholesterol trafficking, and mitochondrial DNA maintenance. It directly interacts with StAR to channel cholesterol to CYP11A1 for steroidogenesis and recruits DRP1 to mediate fission. By sequestering BAX and BAK, ATAD3A also restrains the intrinsic apoptosis pathway. Its expression is controlled by upstream regulators including PGC-1??, NRF1, and SF-1, integrating mitochondrial biogenesis with metabolic and hormonal signals, while cAMP signaling provides further modulation in steroidogenic contexts.
In HEK293T cells, ATAD3A knockout disrupts cristae architecture and cholesterol import, impairing StAR-dependent trafficking to the inner membrane. Reduced DRP1 recruitment slows fission, leading to elongated mitochondria observable by TOM20 or MitoTracker staining. Derepression of BAX/BAK sensitizes cells to apoptosis, creating a tractable system for studying mitochondrial checkpoint control. The HEK293T background supports co-expression of exogenous regulators like PGC-1?? or SF-1, enabling dissection of hierarchical control within the ATAD3A network.
This knockout model is suited for high-content imaging of mitochondrial dynamics, radioisotope- or fluorescence-based cholesterol transport assays, and Annexin V/PI flow cytometry to measure apoptotic priming. It also enables mtDNA copy number quantification by qPCR and drug screening for mitochondrial vulnerabilities in cancer. For steroidogenic studies, co-treatment with cAMP analogs combined with ELISA-based hormone detection allows assessment of ATAD3A??s role in hormone biosynthesis. The ATAD3A Knockout HEK293T Polyclonal Cells provide a powerful platform for mitochondrial research. For more details, please contact Ascent Research.
