Gene transfer to the lung is critically needed to treat a number of different disease states. Electroporation across the chest following airway delivery of a DNA solution, termed transthoracic electroporation, overcomes many of the limitations seen with viral and molecular conjugate approaches. It has been shown to be highly efficient, simple, safe, and noninflammatory and results in gene transfer to over 90% of the area of the lung. Up to 50% of the cells in the parenchyma take up and express transgene as do up to 80% of the airway cells. The method has been applied to both small (mouse and rat) and large (pig) animal models, yielding similar results. As such, this method has been used extensively to develop treatments for acute respiratory distress syndrome (ARDS). ARDS is a common, devastating clinical syndromes that has up to 30% mortality with the current standard of care. ARDS results from the accumulation and inability to clear pulmonary edema, due to injury of the alveolar epithelium and pulmonary endothelium. Gene therapy may provide treatment of these diseases by restoring alveolar fluid clearance and barrier function. It has been shown that electroporation-mediated gene transfer of several different ion channels can increase fluid clearance from the healthy lung, but only gene transfer of proteins that regulate barrier function can lead to improvements in a previously injured lung. The β1 subunit of the Na+, K+-ATPase has been shown to act through the kinase MRCKα to upregulate cortical actin assembly, leading to increases in tight and adherens junction abundance and activity. Electroporation-mediated gene transfer of either the Na+, K+-ATPase β1 subunit or MRCKα leads to decreased lung permeability, upregulated barrier function, and reduced inflammation. These treatment approaches provide benefit in both mouse models and in a pig model that accurately duplicates the complex inflammatory and hemodynamic response that occurs in humans with ARDS. The success of this electroporation-mediated gene therapy in such a stringent model clearly demonstrates the robust nature of this approach and warrants preclinical testing as a prelude to use in critically ill patients.