Introduction: Multiple organ failure is the commonest cause of death in septic patients. Objectives: This study was undertaken in an attempt to elucidate the functional importance of DNA -dependent protein kinase catalytic subunit (DNA-PKcs) on mitochondrial dysfunction associated with the development and progression of sepsis-related multiple organ dysfunction syndrome (MODS). Methods: Cardiomyocyte-specific DNA-PKcs knockout (DNA-PKcsCKO) mice, liver-specific DNA-PKcs knockout (DNA-PKcsLKO) mice, and kidney tubular cell-specific DNA-PKcs knockout (DNA-PKcsTKO) mice were used to generate an LPS-induced sepsis model. Echocardiography, serum biochemistry, and tissue microscopy were used to analyze organ damage and morphological changes induced by sepsis. Mitochondrial function and dynamics were determined by qPCR, western blotting, ELISA, and mt-Keima and immunofluorescence assays following siRNA-mediated DNA-PKCs knockdown in cardiomy-ocytes, hepatocytes, and kidney tubular cells.Results: DNA-PKcs deletion attenuated sepsis-mediated myocardial damage through improving mito-chondrial metabolism. Loss of DNA-PKcs protected the liver against sepsis through inhibition of mito-chondrial oxidative damage and apoptosis. DNA-PKcs deficiency sustained kidney function upon LPS stress through normalization of mitochondrial fission/fusion events, mitophagy, and biogenesis.Conclusion: We conclude that strategies targeting DNA-PKcs expression or activity may be valuable ther-apeutic options to prevent or reduce mitochondrial dysfunction and organ damage associated with sepsis-induced MODS.(c) 2022 The Authors. Published by Elsevier B.V. on behalf of Cairo University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).