Gene therapy represents a potentially useful approach for the treatment of diseases refractory to conventional therapies. Various preclinical and clinical strategies have been explored for treatment of gynaecological diseases. Given the most severe unmet clinical need, much of the work has been performed with gynaecological cancers and ovarian cancer in particular. Although the safety of many treatment strategies has been demonstrated in early phase clinical trials, efficacy has been mostly limited heretofore. Major challenges include improving the vectors used with the aim of more effective and selective delivery. In addition, effective penetration into and spreading within advanced and complex tumour masses and metastases remains challenging. This review focuses on existing and developmental gene transfer applications for gynaecological diseases.