ISOC2 Knockout HEK293 Cell Line

The ISOC2 Knockout HEK293 Cell Line is a CRISPR/Cas9-edited human cell line providing a loss-of-function model for the ISOC2 gene. Targeted ISOC2 disruption enables investigation of its roles in mitochondrial metabolism, pyrimidine biosynthesis, and cell proliferation within a well-characterized HEK293 background. This knockout cell line offers a reliable system for studying ISOC2-dependent pathways without requiring transient suppression methods.

HEK293 cells are human embryonic kidney epithelial cells transformed with sheared adenovirus 5 DNA, resulting in a hypotriploid karyotype and high transfection efficiency. Their adherent growth, partial Ad5 genome, and robust protein expression capacity make them a preferred system for heterologous gene expression, viral production, and functional genomics. This background supports consistent experimental outcomes in knockout studies focused on mitochondrial and metabolic gene functions.

ISOC2 encodes a mitochondrial protein with predicted isochorismatase activity and is regulated by E2F transcription factors and MYC. Within the mitochondria, ISOC2 interacts with matrix proteins and nucleoside metabolism enzymes such as DHODH and UMP synthase, linking pyrimidine biosynthesis to apoptosis signaling. ISOC2 knockout disrupts mitochondrial membrane integrity, promoting cytochrome c release and caspase-9 activation, while altering the balance of BCL2 family members BAX and BCL2. Consequently, pyrimidine biosynthesis is impaired, and cells become more susceptible to apoptotic stimuli, offering a platform to study metabolic regulation of cell death.

In HEK293 cells, ISOC2 knockout markedly affects mitochondrial metabolic processes that drive proliferation and survival. The metabolic activity of HEK293 cells amplifies the phenotypes resulting from ISOC2 loss, including reduced proliferation and increased apoptosis. This cell line thus serves as a valuable model for dissecting how mitochondrial dysfunction influences pyrimidine biosynthesis, cell cycle regulation, and apoptotic sensitivity in a human epithelial context. It enables direct interrogation of ISOC2 function using a wide array of biochemical and cell-based approaches.

The ISOC2 Knockout HEK293 Cell Line supports diverse applications in mitochondrial metabolism, cancer cell proliferation, and apoptosis research. With ISOC2 overexpression reported in hepatocellular carcinoma and non-small cell lung cancer, this model is ideal for drug screening targeting mitochondrial functions and for mechanistic studies of cell cycle dysregulation. Key assays include Western blotting for BCL2, BAX, and cleaved caspase-9; RT-qPCR for metabolic genes; MTT or CCK-8 proliferation assays; flow cytometry with TMRE or JC-1 for mitochondrial membrane potential; caspase activity measurements; metabolomic profiling; and RNA-seq. For further information, contact Ascent Research.