Journal Article

PUBDB-2025-05098

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Lipid Monolayers Adsorbing mRNA as Models for mRNA Enclosed in Lipid Nanoparticles for Transfection

Grava, M.Extern* ; Weber, K.Extern* ; Reed, J.Extern* ; Weber, B.Extern*EMBL* ; Shen, C.Extern*DESY* ; Haas, H. (Corresponding author)Extern*EMBL* ; Schneck, E. (Corresponding author)Extern*

2025
ACS Publ. Washington, DC

This record in other databases:    

Please use a persistent id in citations: doi:10.1021/acs.langmuir.5c04076  doi:10.3204/PUBDB-2025-05098

Abstract: In lipid-based mRNA pharmaceuticals applicablefor vaccination, cancer therapy, and other types of therapeutics,negatively charged RNA is embedded in nanoparticles containingpositively charged or ionizable lipids. The local cohesion at thelipid-RNA interface plays an important role in the stability andbiological activity of the resulting nanoparticles. Ions and otherbuffer components in the RNA’s immediate surroundings mayaffect RNA stability against hydrolysis and its binding strength tothe oppositely charged lipid layers, which is relevant for releaseafter cellular uptake and endosomal processing. Here, we useLangmuir monolayers at the air−water interface as well-definedexperimental models to study the local molecular organization ofmRNA adsorbed to lipid layers. The binding of mRNA(approximately 3900 nucleotides) in the presence of monovalent and divalent ions from the aqueous phase to monolayersconsisting of cationic transfection lipids and zwitterionic phospholipids is investigated with synchrotron-based X-ray fluorescenceand scattering techniques. The experiments provide detailed insights into the structure of the adsorbed layers as well as the fractionsof all molecular moieties contributing to the interfacial electrostatic balance, specifically phosphates from RNA, phosphates fromphospholipids in the monolayer, and elemental counterions such as chloride, potassium, and calcium. The mRNA forms discrete, 1−2 nm thick, electron-dense layers tightly bound to the lipid membrane, where cationic ions accumulate at the interface together withthe mRNA. This leads to a 1.5−3-fold excess of anionic nucleotides relative to cationic lipids at the interface, which is morepronounced in the presence of divalent compared to monovalent cations. The anions that compensate for the lipid charge in theabsence of RNA get displaced from the interface upon RNA adsorption. The quantitative information about local interfacial moietiesobtained here may constitute a valuable basis for the evaluation of quality aspects of nanoparticles comprising mRNA in the bulkphase.

Note: ErUM-Pro 05K19FK2

---

Contributing Institute(s):

1. PETRA-D (FS-PETRA-D)
2. DOOR-User (DOOR ; HAS-User)

Research Program(s):

1. 633 - Life Sciences – Building Blocks of Life: Structure and Function (POF4-633) (POF4-633)
2. 6G3 - PETRA III (DESY) (POF4-6G3) (POF4-6G3)
3. FS-Proposal: I-20220194 (I-20220194) (I-20220194)
4. FS-Proposal: I-20220952 (I-20220952) (I-20220952)
5. FS-Proposal: I-20230480 (I-20230480) (I-20230480)

Experiment(s):

1. PETRA Beamline P08 (PETRA III)

Appears in the scientific report 2025

---

Database coverage:
; ; Clarivate Analytics Master Journal List ; Current Contents - Physical, Chemical and Earth Sciences ; Ebsco Academic Search ; Essential Science Indicators ; IF < 5 ; JCR ; Nationallizenz ; SCOPUS ; Science Citation Index Expanded ; Web of Science Core Collection

---