锌指核酸内切酶:基因操作的有力工具

Zinc Finger Nuclease,A Powerful Tool for Gene Manipulation

针对动植物进行基因靶向操作的技术,是解析基因功能、研究疾病,以及农业经济生产中一个有用的工具。至今,基因靶向操作主要是通过在胚胎干细胞(embryonic stem cell,ES cell)中进行同源重组或者是体细胞核转移的方法进行,但同源重组方法由于需要ES细胞而被限制在个别物种,而核转移方法存在核去分化、效率低、成本高的缺陷。近几年,一种基于锌指核酸内切酶(zinc-finger nuclease,ZFN)基因靶向修饰的新技术被应用于包括植物、果蝇、爪蟾、斑马鱼和大鼠等不同物种的基因操作。通过胚胎注射ZFN的质粒或是mRNA可以有效地定靶并迅速地在内源基因上引起可遗传的突变。ZFN介导基因靶向敲除的可行性,使得那些无法获得ES细胞和克隆技术支持的物种的基因靶向修饰成为可能。

The generation of genetically modified animals or plants with gene-targeting is a powerful tool to analyze gene function, study disease and produce agriculture organisms of economical interest.Until recently, the gene targeted manipulations of animals accomplished by homologous recombination （HR） in embryonic stem （ES） cells or nuclear transfer of somatic cells have been limited to a few species because of the ES cell availability and the difficulties in dedifferentiation, efficiency, and the cost of somatic nuclear transfer.Recently, a new technology based on the use of zinc-finger nucleases （ZFNs） was developed and used to generate genome engineering organisms such as plants, Drosophila, zebra fish and rats with gene targeting.Microinjection of plasmid or mRNA for ZFNs into embryos allowed rapid, efficient, permanent and heritable disruption of endogenous loci.Therefore, the application of ZFNs is a promising strategy to perform gene targeting in species where ES cells or cloning techniques are not available.