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Background: Infections caused by multidrug-resistant (MDR) Gram-negative bacteria are rising. Bacteriophages are suggested as an alternative treatment option for the treatment of antibiotic-resistant bacteria. Bacteriophages in treatment of Gram-negative bacterial infections is not well investigated in vivo. The aim of this study was to review systematically the studies on bacteriophages against infection caused by Gram-negative bacteria in vivo. Methods: This systematic review was done using electronic databases, including Scopus, PubMed, Google Scholar, and Web of science; the articles published from 1983 to 2018 were investigated. Studies meeting the inclusion criteria were selected, and the data were estimated using a review method. Findings: 1310 articles were indexed from which 380 were selected based on their abstracts. Then, some were excluded including clinical trials and in-vitro studies. Finally, experimental studies (n = 31), that met the inclusion criteria and were published in English, were selected. Conclusion: This review showed that bacteriophages are an effective treatment against n-vivo Gram-negative bacteria infections even be used orally, topically, or subcutaneously injected.