This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 cell line, with disruption of the ATAD1 gene (ATPase family AAA domain containing 1). As a polyclonal preparation, it provides a heterogeneous pool of cells carrying diverse loss-of-function mutations, enabling robust functional studies without clonal selection. The model is intended for researchers investigating ATAD1-mediated mitochondrial biology in a human liver cancer background.
The SK-HEP-1 cell line originates from the ascites of a 52-year-old male with liver adenocarcinoma. Although initially misclassified as endothelial, these cells are now known to be poorly differentiated hepatocellular carcinoma with an epithelial-like adherent morphology. SK-HEP-1 is extensively used in hepatocarcinogenesis, tumor progression, and metastasis research, retaining aggressive features such as high proliferation and metastatic capacity.
ATAD1 encodes a mitochondrial AAA+ ATPase that extracts mislocalized tail-anchored (TA) proteins from the outer mitochondrial membrane, a critical function for mitochondrial proteostasis. It is regulated by transcription factor NRF1 (NFE2L1) and influenced by mitochondrial membrane potential and import stress. Core substrates include PINK1, Fis1, and Gem1; their accumulation upon ATAD1 loss disrupts quality control and alters apoptosis through regulators like BAX and BAK. ATAD1 interacts with the TOM complex (e.g., TOMM70) and coordinates with the E3 ligase MARCH5 and the ubiquitin-proteasome system to degrade errant TA proteins.
In hepatocellular carcinoma, mitochondrial dysfunction is tightly linked to chemoresistance, metabolic reprogramming, and apoptotic evasion. This polyclonal ATAD1 knockout model enables examination of how TA protein mislocalization affects cellular fitness, stress responses, and apoptotic thresholds in a poorly differentiated liver cancer setting. Notably, ATAD1 dysregulation has been observed in some cancers, making this tool valuable for clarifying whether ATAD1 functions as a tumor suppressor or survival factor in hepatic tumors.
The product supports diverse applications including mitochondrial isolation coupled with proteinase K protection assays and Western blotting for PINK1, Fis1, and Gem1. Apoptosis can be measured by Annexin V/caspase assays, with complementary cell viability (MTT) and ROS analyses. Seahorse respirometry enables metabolic profiling, and co-immunoprecipitation probes ATAD1?CTA protein interactions. Additionally, the polyclonal cells are suitable for RNA-seq and high-throughput screens to discover mitochondrial quality control modulators. For further information, please contact Ascent Research.
