Nonviral gene delivery by tetraamino fullerene
Nonviral gene delivery by tetraamino fullerene
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DOI：
10.1021/mp050068r
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发表时间：
2006-03-01
影响因子：
4.9
通讯作者：
Nakamura, Eiichi
中科院分区：
文献类型：
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作者：
Isobe, Hiroyuki;Nakanishi, Waka;Nakamura, Eiichi
带有两个二氨基侧链的富勒烯衍生物与大小为4或40 kbp的质粒载体DNA结合，在孵育时将其递送至哺乳动物细胞，并导致编码基因的瞬时或稳定表达。对各种富勒烯化合物对核酸酶的DNA结合和保护性质的初步物理化学研究使我们确定四氨基富勒烯作为探测富勒烯介导的基因递送到哺乳动物细胞的新概念的理想候选物。利用绿色荧光蛋白和荧光素酶报告基因对COS-1细胞进行瞬时和稳定转染的研究表明，与传统的脂质体转染方法相比，富勒烯转染具有许多有用的特性，包括更高的稳定转染效率和转染融合细胞的能力。化学和生物学研究表明，富勒烯/DNA复合物的细胞摄取通过内吞作用机制发生，并且由内体内化的DNA受到富勒烯的保护，免受酶消化。富勒烯/DNA复合物的刚性可能在富勒烯方法的成功中发挥一定作用。
A fullerene derivative bearing two diamino side chains binds to a plasmid vector DNA, either 4 or 40 kbp in size, delivers it to mammalian cells on incubation, and leads to expression of the encoded gene either transiently or stably. The initial physicochemical investigations upon DNA-binding and protective properties of various fullerene compounds against nuclease led us to identify the tetraamino fullerene as an ideal candidate to probe the new concept of fullerene-mediated gene delivery to mammalian cells. Studies on transient and stable transfection of COS-1 cells using green fluorescent protein and luciferase reporter genes revealed several useful properties of the fullerene transfection as compared with the conventional lipid-based transfection method, including much higher efficiency of stable transfection and ability to transfect confluent cells. Chemical and biological studies suggested that the cell uptake of the fullerene/DNA complex takes place by an endocytosis mechanism and that the DNA internalized by endosomes is protected by the fullerene against enzymatic digestion. The stiffness of the fullerene/DNA complex may play some role in the success of the fullerene method.