The SIRT3 Knockout AC16 Cell Line is a CRISPR/Cas9-edited knockout cell line that disrupts the SIRT3 gene, providing a stable loss-of-function model for investigating mitochondrial biology in a human cardiomyocyte context. This cell line enables researchers to dissect SIRT3-dependent pathways in metabolism, oxidative stress, and energy homeostasis without interference from wild-type activity, making it a valuable tool for mechanistic and translational studies.
The AC16 host cell line is an immortalized human adult ventricular cardiomyocyte line generated by fusion with SV40-transformed human fibroblasts. AC16 cells maintain key cardiomyocyte characteristics, including expression of cardiac-specific markers and contractile ability, and are widely employed as a model for studying human cardiac function, pathology, and pharmacological responses. Their mature ventricular phenotype is particularly relevant for modeling adult-onset cardiovascular diseases such as heart failure, hypertrophy, and ischemic injury.
SIRT3 encodes a mitochondrial NAD+-dependent protein deacetylase that serves as a central regulator of metabolic adaptation and antioxidant defense. Within the mitochondrial matrix, SIRT3 deacetylates and activates critical enzymes including isocitrate dehydrogenase 2 (IDH2), superoxide dismutase 2 (SOD2), long-chain acyl-CoA dehydrogenase (LCAD), and glutamate dehydrogenase (GDH), thereby enhancing oxidative phosphorylation, fatty acid ??-oxidation, and reactive oxygen species detoxification. SIRT3 function is controlled by upstream regulators such as PGC-1??, NRF1, NRF2, and NAD+ availability, and it interacts with mitochondrial complex I, OPA1, and sirtuins SIRT4 and SIRT5. Downstream deacetylation of FOXO3a and cyclophilin D further links SIRT3 to autophagy and mitochondrial permeability transition, positioning it at the nexus of mitochondrial quality control.
In cardiomyocytes, SIRT3 is essential for sustaining the high metabolic demands of the heart. Disruption of SIRT3 in AC16 cells leads to impaired mitochondrial respiration, elevated reactive oxygen species, and altered substrate utilization, phenocopying features of cardiac stress and disease. This knockout model is therefore highly relevant for studying the molecular mechanisms underlying diabetic cardiomyopathy, cardiac hypertrophy, ischemia-reperfusion injury, and age-related cardiac dysfunction, allowing precise delineation of SIRT3??s contribution to cardiac pathophysiology.
The SIRT3 Knockout AC16 Cell Line supports a broad spectrum of research applications, including cardiovascular disease modeling, mitochondrial dysfunction studies, and metabolic stress assays. Researchers can employ this line for Seahorse-based respiration analysis, ATP quantification, ROS measurement, apoptosis evaluation via TUNEL or caspase activity, and drug screening for cardioprotective agents. Standard analytical techniques such as western blotting, RT-qPCR, immunofluorescence, and metabolomics are fully compatible. Additionally, it serves as a platform for aging and longevity investigations. For further information, please contact Ascent Research.
