Tabriz University of Medical Sciences Advanced Pharmaceutical Bulletin 2228-5881 15 3 2025 09 25 Genosomes: An Introspection into Transfection, Future Perspectives and Applications 482 498 10.34172/apb.025.43191 EN Abhirami Subramony https://orcid.org/0009-0005-9049-2081 Divyatha Raj Fiza Fairooz Aparna Venugopal Aparna Rajesh Sreeja C Nair https://orcid.org/0000-0001-9861-0461 REVIEW 10.34172/apb.025.43191 2024 06 22 2025 08 27 The advent of advanced gene delivery platforms has transformed the precision targeting of therapeutic nucleic acids, such as miRNA, siRNA, and pDNA, for the treatment of genetic and acquired diseases, including cystic fibrosis, malignancies, sickle cell anaemia, and β-thalassemia. Although viral vectors have traditionally dominated this field, non-viral systems, particularly genosomes (cationic lipid-based nanocarriers or lipoplexes), have emerged as promising alternatives due to their enhanced biosafety, lower immunogenic potential, and manufacturability. These nanostructured systems facilitate efficient nucleic acid condensation, protect against enzymatic degradation, and enhance cellular uptake and endosomal escape. Further refinements, including PEGylation, incorporation of helper lipids, and stimuli-responsive formulations, have significantly improved transfection efficiency and tissue-specific delivery. Notable clinical advancements, such as RNA-lipoplexes in cancer immunotherapy and multifunctional envelope-type nanodevices (MEND), highlight their therapeutic potential. This review provides a critical analysis of genosome design strategies, formulation techniques, intracellular trafficking mechanisms, clinical applications, patented innovations, and future prospects to advance genosome-mediated gene therapy.