Many potent biologics are limited by their short half-lives, necessitating frequent administration, which can be burdensome for patients and economically costly. Granulocyte-colony stimulating factor (G-CSF), a critical treatment for severe chronic neutropenia (SCN), known under the generic drug name filgrastim, exemplifies this challenge. SCN patients often require daily doses of G-CSF, which can adversely affect compliance and highlights the need for improved therapeutic strategies. Intramuscular electroporation (IM-EP) of DNA medicines has emerged as a promising solution by facilitating continuous and long-lasting production of the encoded therapeutic protein. Here, we employ this technology for a DNA-based delivery of G-CSF. The data from mouse models demonstrate that a single administration leads to an improved pharmacokinetic profile, durable gene expression beyond one year, and efficacy in a neutropenia disease model. Furthermore, results from rabbit models validate the scalability of this approach and confirm that muscle-derived G-CSF specifically increases neutrophil count without affecting other bone marrow-derived cell types. These findings underscore the potential of IM-EP of DNA medicines as an effective method to overcome the limitations posed by the short half-life of some biologics, offering a more patient-friendly and cost-effective therapeutic approach.