In vivo delivery of large RNA molecules has significant implications for novel gene therapy, biologics delivery, and vaccine applications. We have developed cationic nanolipoprotein particles (NLPs) to enhance the complexation and delivery of large self-amplifying mRNAs (replicons) in vivo. NLPs are high-density lipoprotein (HDL) mimetics, comprised of a discoidal lipid bilayer stabilized by apolipoproteins that are readily functionalized to provide a versatile delivery platform. Herein, we systematically screened NLP assembly with a wide range of lipidic and apolipoprotein constituents, using biophysical metrics to identify lead candidates for in vivo RNA delivery. NLPs formulated with cationic lipids successfully complexed with RNA replicons encoding luciferase, provided measurable protection from RNase degradation, and promoted replicon in vivo expression. The NLP complexation of the replicon and in vivo transfection efficiency were further enhanced by modulating the type and percentage of cationic lipid, the ratio of cationic NLP to replicon, and by incorporating additive molecules.

中文翻译：

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阳离子HDL模拟物可增强体内自我复制mRNA的传递。

大分子RNA的体内递送对新型基因治疗，生物制剂递送和疫苗应用具有重要意义。我们已经开发出阳离子纳米脂蛋白颗粒（NLP），以增强体内大型自扩增mRNA（复制子）的络合和传递。NLP是高密度脂蛋白（HDL）模拟物，由通过载脂蛋白稳定的盘状脂质双层组成，可以容易地功能化以提供通用的递送平台。在本文中，我们使用生物物理指标来识别体内RNA输送的潜在候选对象，系统地筛选了具有广泛脂质和载脂蛋白成分的NLP组装。由阳离子脂质配制的NLP与编码萤光素酶的RNA复制子成功复合，可针对RNase降解提供可测量的保护，并促进复制子的体内表达。通过调节阳离子脂质的类型和百分比，阳离子NLP与复制子的比例以及掺入添加剂分子，可进一步增强复制子的NLP络合和体内转染效率。