Baculovirus-mediated gene transfer into pancreatic islet cells
Baculovirus-mediated gene transfer into pancreatic islet cells
复制标题
DOI：
10.2337/diabetes.49.12.1986
复制
发表时间：
2000-12-01
期刊：
影响因子：
7.7
通讯作者：
Philipson, LH
中科院分区：
文献类型：
--
作者：
Ma, L;Tamarina, N;Philipson, LH
杆状病毒转导是一种基因转移方法，其使用蛾细胞病毒用于培养中的哺乳动物细胞，这导致高水平的延长表达。在这里，我证明了重组杆状病毒可以作为有效的基因转移载体，用于将哺乳动物启动子驱动的外源基因递送到人和小鼠胰岛细胞中。现有的方法，如各种转染和电穿孔技术，要么效率低，要么造成广泛的膜损伤。病毒载体已成为哺乳动物细胞中基因递送和表达的重要工具，但存在一些缺点，例如构建时间长和病毒基因的表达。苜蓿银纹夜蛾多核型多角体病毒（Autographa californica multiple nuclear polyhedrosis virus）作为外源基因在昆虫细胞中表达的载体被广泛使用，并且最近在一些哺乳动物细胞中也被广泛使用。在巨细胞病毒启动子盒中使用几种绿色荧光蛋白和LacZ表达构建体，我在原代人和小鼠胰岛细胞中获得了有效的基因表达。杆状病毒感染后，葡萄糖刺激的细胞内游离钙反应没有受损。病毒的安全性以及构建和繁殖的相对容易性使得杆状病毒系统成为促进外源基因转移的有用工具。
Baculovirus transduction is a gene transfer method that uses a moth cell virus for mammalian cells in culture, which results in a high-level prolonged expression. Here me demonstrate that recombinant baculoviruses can serve as efficient gene transfer vehicles for delivering foreign genes driven by mammalian promoters into human and mouse pancreatic islet cells. Existing methods, such as various transfection and electroporation techniques, either suffer from low efficiency or cause extensive membrane damage. Viral vectors have emerged as an important tool for gene delivery and expression in mammalian cells but suffer from several drawbacks, such as lengthy construction time and expression of viral genes. The baculovirus Autographa californica multiple nuclear polyhedrosis virus is widely used as a vector for expression of foreign genes in insect cells and, more recently, in some mammalian cells. Using several green fluorescent protein- and LacZ-expressing constructs in a cytomegalovirus promoter cassette, me obtained efficient gene expression in primary human and mouse islet cells. There was no impairment of glucose-stimulated intracellular free calcium responses after baculovirus infection. The safety and the relative ease of construction and propagation of the virus makes the baculovirus system a useful tool for facilitating the transfer of foreign genes.