Multi-layered nanoparticles for penetrating the endosome and nuclear membrane via a step-wise membrane fusion process
Multi-layered nanoparticles for penetrating the endosome and nuclear membrane via a step-wise membrane fusion process
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DOI：
10.1016/j.biomaterials.2009.02.009
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发表时间：
2009-05-01
期刊：
影响因子：
14
通讯作者：
Harashima, Hideyoshi
中科院分区：
文献类型：
--
作者：
Akita, Hidetaka;Kudo, Asako;Harashima, Hideyoshi
将DNA有效靶向细胞核是有效基因治疗的先决条件。基因递送载体必须穿透质膜和DNA不可渗透的双层膜核被膜，并以准备转录的形式存款其DNA货物。在这里，我们介绍了一个概念，克服细胞内膜障碍，涉及逐步膜融合。为了实现这一目标，开发了一种纳米技术，可以创建多层纳米颗粒，我们将其称为四层多功能包膜型纳米器件（T-MEND）。T-MEND的关键结构元素是一个DNA-聚阳离子凝聚核心，由两个核膜-融合的内包膜和两个溶酶体-融合的外包膜包裹，它们以逐步的方式脱落。通过双层核膜结构的逐步融合进行核递送需要双层膜结构。通过荧光共振能量转移（FRET）的光谱成像的供体和受体荧光团，已双重标记的脂质体表面之间的细胞内膜融合的内体和核膜进行了验证。用最少数量的核融合脂质包膜（即，2)对促进转录至关重要。因此，T-MEND在非分裂细胞中实现了显著水平的转基因表达。(C)2009爱思唯尔有限公司保留所有权利。
Efficient targeting of DNA to the nucleus is a prerequisite for effective gene therapy. The gene-delivery vehicle must penetrate through the plasma membrane, and the DNA-impermeable double-membraned nuclear envelope, and deposit its DNA cargo in a form ready for transcription. Here we introduce a concept for overcoming intracellular membrane barriers that involves step-wise membrane fusion. To achieve this, a nanotechnology was developed that creates a multi-layered nanoparticle, which we refer to as a Tetra-lamellar Multi-functional Envelope-type Nano Device (T-MEND). The critical structural elements of the T-MEND are a DNA-polycation condensed core coated with two nuclear membrane-fusogenic inner envelopes and two enclosome-fusogenic outer envelopes, which are shed in stepwise fashion. A double-lamellar membrane structure is required for nuclear delivery via the stepwise fusion of double layered nuclear membrane structure. Intracellular membrane fusions to endosomes and nuclear membranes were verified by spectral imaging of fluorescence resonance energy transfer (FRET) between donor and acceptor fluorophores that had been dually labeled on the liposome surface. Coating the core with the minimum number of nucleus-fusogenic lipid envelopes (i.e., 2) is essential to facilitate transcription. As a result, the T-MEND achieves dramatic levels of transgene expression in non-dividing cells. (C) 2009 Elsevier Ltd. All rights reserved.