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LNPs based on 1,2\u2010dioleoyl\u20103\u2010trimethylammonium propane (DOTAP) have been widely used in preclinical and clinical settings. A novel non\u2010viral gene delivery system called LNP3 was previously developed, which was composed of DOTAP, 1,2\u2010dioleoyl\u2010sn\u2010glycero\u20103\u2010phosphoethanolamine (DOPE), and cholesterol. One of the helper lipids in this carrier was DOPE, which belongs to phospholipids. Given that substituting DOPE with non\u2010phospholipids as helper lipids can increase the delivery efficiency of some LNPs, this study aimed to examine whether non\u2010phospholipids can be formulated with DOTAP as helper lipids. It was found that monoglycerides with C14:0, C16:0, C18:0, C18:1, and C18:2 mediated mRNA transfection, and the transfection efficiency varied between C18:0, C18:1, and C18:2. Furthermore, substituting of the glycerol with other moieties such as the cholesterol or the ethanolamine similarly mediated mRNA transfection. The introduction of cholesterol can further improve the transfection capacity of some DOTAP\u2010based LNPs. One of the best\u2010performing formulations, LNP3\u2010MO, was used to mediate luciferase\u2010mRNA expression in vivo, and the luminescence signal was found to be mainly enriched in the lung and spleen. In addition, the level of SARS\u2010CoV\u20102 spike antibody in the serum increased after three doses of LNP3\u2010MO mediated SARS\u2010CoV\u20102 spike mRNA. Altogether, this study demonstrates that non\u2010phospholipids are promising helper lipids that can be formulated with DOTAP to facilitate efficient delivery of mRNAs in vitro and in vivo with organ\u2010specific targeting.<\/jats:p>","DOI":"10.1002\/biot.202300123","type":"journal-article","created":{"date-parts":[[2023,8,7]],"date-time":"2023-08-07T03:06:46Z","timestamp":1691377606000},"source":"Crossref","is-referenced-by-count":15,"title":["Exploration of mRNA nanoparticles based on DOTAP through optimization of the helper lipids"],"prefix":"10.1002","volume":"18","author":[{"given":"Xueni","family":"Ma","sequence":"first","affiliation":[{"name":"Key Laboratory of Digestive Diseases Lanzhou University Second Hospital  Lanzhou China"},{"name":"The Second Clinical Medical College Lanzhou University  Lanzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fanqi","family":"Wu","sequence":"additional","affiliation":[{"name":"The Second Clinical Medical College 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