Insertion of high molecular weight messenger RNA (mRNA) into lyotropic lipid phases as model systems for controlled release formulations for the mRNA was investigated. Low fractions of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) were used as an anchor to load the mRNA into a lamellar lipid matrix. Dispersions of zwitterionic lipid in the aqueous phase in the presence of increasing fractions of mRNA and cationic lipid were prepared, and the molecular organization was investigated as a function of mRNA and cationic lipid fraction. Insertion of both cationic lipid and mRNA was clearly proven from the physicochemical characteristics. The d-spacing of the lipid bilayers, as determined by small-angle X-ray scattering (SAXS) measurements, responded sensitively to the amount of inserted DOTAP and mRNA. A concise model of the insertion of the mRNA in the lipid matrices was derived, indicating that the mRNA was accommodated in the aqueous slab between lipid bilayers. Depending on the DOTAP and mRNA fraction, a different excess of water was present in this slab. Results from further physicochemical characterization, including determination of free and bound mRNA, zeta potential, and calorimetry data, were in line with this assumption. The structure of these concentrated lipid/mRNA preparations was maintained upon dilution. The functionality of the inserted mRNA was proven by cell culture experiments using C2C12 murine myoblast cells with the luciferase-encoding mRNA. The described lipid phases as carriers for the mRNA may be applicable for different routes of local administration, where control of the release kinetics and the form of the released mRNA (bound or free) is required.

中文翻译：

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mRNA在层状脂质基质中的肠胃外给药

研究了将高分子量信使RNA（mRNA）插入溶致脂质相中作为mRNA控释制剂的模型系统。低级分的1,2-二油酰基-3-三甲基铵-丙烷（DOTAP）被用作将mRNA加载到层状脂质基质中的锚点。在存在增加的mRNA和阳离子脂质组分的情况下，制备了两性离子脂质在水相中的分散体，并研究了分子结构与mRNA和阳离子脂质组分的关系。阳离子脂质和mRNA的插入均已从理化特性中得到明确证明。该d通过小角度X射线散射（SAXS）测量确定的脂双层的-spacing对插入的DOTAP和mRNA的量敏感。得到了在脂质基质中mRNA插入的简明模型，表明mRNA被容纳在脂质双层之间的水性平板中。根据DOTAP和mRNA的比例，此平板中存在不同程度的过量水。进一步的物理化学表征（包括测定游离和结合的mRNA，ζ电位和量热数据）的结果与该假设相符。这些浓缩的脂质/ mRNA制品的结构在稀释后得以保持。通过使用带有荧光素酶编码mRNA的C2C12鼠成肌细胞进行细胞培养实验，证明了插入的mRNA的功能。