Description

The Sucla2 Knockout NIH 3T3 Cell Line is a validated CRISPR/Cas9-edited knockout cell line that provides targeted disruption of the Sucla2 gene in a mouse embryonic fibroblast background. This gene-edited model enables investigation of the succinyl-CoA ligase beta subunit without relying on specific mutation information, offering a rigorous loss-of-function platform for metabolic studies.

NIH 3T3 cells are immortalized murine embryonic fibroblasts that are widely adopted in gene editing, signal transduction, and cell biology research owing to their robust growth and ease of manipulation. Their fibroblast origin offers a physiologically relevant system for dissecting mitochondrial function and energy metabolism, making them a suitable host for this knockout.

Sucla2 encodes the beta subunit of succinyl-CoA synthetase (SCS), which together with the alpha subunit SUCLG1 catalyzes the reversible conversion of succinyl-CoA and ADP to succinate and ATP. This reaction represents a critical substrate-level phosphorylation step within the TCA cycle. Sucla2 expression is transcriptionally regulated by PPARGC1A and PPARA, which respond to nutrient availability and AMPK signaling. The enzyme interacts with SUCLG2 and mitochondrial matrix chaperones to form functional complexes. Its activity generates succinate, which feeds into complex II of the electron transport chain, and directly contributes to the mitochondrial ATP pool.

Knockout of Sucla2 in NIH 3T3 cells disrupts TCA cycle flux and substrate-level phosphorylation, leading to reduced ATP synthesis and altered metabolic homeostasis. This deficiency models succinyl-CoA ligase dysfunction found in mitochondrial DNA depletion syndrome and metabolic encephalomyopathy, providing a relevant cellular platform to dissect mitochondrial stress responses and metabolic reprogramming under various nutrient conditions.

This cell line is suitable for diverse experimental applications, including TCA cycle metabolite profiling by LC-MS, assessment of mitochondrial respiration using Seahorse technology, measurement of succinyl-CoA ligase enzymatic activity, RT-qPCR analysis of metabolic gene expression, cellular ATP quantification, and cell proliferation or apoptosis assays following metabolic perturbation. These tools enable detailed investigation of the role of Sucla2 in mitochondrial energy metabolism and disease pathogenesis. For further information, please contact Ascent Research.