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In this study, we use giant unilamellar vesicles (GUVs) mimicking the endosomal membrane to directly visualize the interaction and membrane disruptions caused by two model LNPs constituted by two different ionizable lipids: LP01 and BiP-20 (a Branched ionizable Phospholipid) that are both benchmark ionizable lipids for in vivo gene editing in liver. Despite identical TNS-measured apparent pKa (6.6) for both lipids in our conditions, LP01 LNP showed maximal membrane engagement at pH 5.8, while BiP20 LNP required pH 5.1, demonstrating that bulk pKa alone cannot predict the pH at which functional membrane interaction occurs. Using confocal microscopy, we show that the LNP engagement with the endosomal membrane proceeds through first electrostatic attraction, clustering, membrane disruption by forming highly curved morphologies, and finally leading to the rupture of the whole endosomal mimicking membrane. Notably, the mRNA cargo is released only upon complete lysis of the membrane. This indicates that the membrane destabilization and cargo release are mechanistically distinct steps. These findings establish a cell-free platform for dissecting individual sub-steps of endosomal escape.<\/jats:p>","DOI":"10.64898\/2026.09.11.751062","type":"posted-content","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T05:35:16Z","timestamp":1789364116000},"source":"Crossref","is-referenced-by-count":0,"title":["Lipid nanoparticle induced model endosomal membrane disruption"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3870-230X","authenticated-orcid":false,"given":"Shreya","family":"Pramanik","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Portland, OR, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aishwarya","family":"Vasudevan","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Portland, OR, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenghua","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Portland, OR, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Archana","family":"Karole","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Portland, OR, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathieu","family":"Berchel","sequence":"additional","affiliation":[{"name":"Univ. Brest, CNRS, CEMCA, UMR 6521, Brest, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul-Alain","family":"Jaffr\u00e8s","sequence":"additional","affiliation":[{"name":"Univ. 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