The ATAD3A Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatic adenocarcinoma cell line SK-HEP-1. This product provides a heterogeneous loss-of-function model for ATAD3A, generated through CRISPR/Cas9-mediated gene disruption. The polyclonal format avoids clonal selection artifacts, making it suitable for pooled functional genomics screens and population-level phenotypic analyses.
SK-HEP-1 is a widely used liver cancer cell model originally isolated from the hepatic adenocarcinoma of a 52-year-old male. The cell line exhibits a hypertriploid karyotype, wild-type p53, and an activating KRAS G12V mutation. These genetic features underpin its utility in studying oncogenic signaling, metastatic behavior, and mechanisms of drug resistance in hepatocellular carcinoma.
ATAD3A encodes a mitochondrial AAA ATPase that functions at endoplasmic reticulum (ER)?Cmitochondria contact sites, where it interacts with VAPB and PTPIP51 to regulate cholesterol trafficking and mitochondrial morphology. It is activated by p53 and mTORC1 signaling, while ER stress inducers can also modulate its expression. Downstream, ATAD3A influences apoptotic thresholds through BAX/BAK regulation, controls mitochondrial fission by interacting with DRP1, and affects steroidogenesis via STAR. Its loss disrupts the ATAD3A?CVAPB?CPTPIP51 tethering complex, leading to mitochondrial fragmentation, impaired oxidative phosphorylation, and enhanced apoptosis sensitivity.
In the SK-HEP-1 background, ATAD3A knockout uncouples mitochondrial function from the pro-survival signaling driven by KRAS G12V and mTOR, making these cells a powerful tool for dissecting the interplay between oncogenic pathways and mitochondrial homeostasis. The wild-type p53 status further permits investigation of p53-mediated apoptotic responses in the absence of ATAD3A. This model is thus especially relevant for understanding how mitochondrial dynamics contribute to hepatocellular carcinoma progression and drug resistance.
Key applications include western blotting for ATAD3A and apoptosis markers, RT-qPCR for mitochondrial gene expression, immunofluorescence to visualize mitochondrial morphology, flow cytometry for apoptosis quantification, and Seahorse metabolic flux analysis. The cells also support migration and invasion assays and drug sensitivity screening, as well as high-throughput synthetic lethal interaction studies. For additional information, please contact Ascent Research.
