Novak Stephen
Corteva Agriscience™, Agriculture Division of DowDuPont™, Indianapolis, IN, USA.
Methods Mol Biol. 2019;1864:295-310. doi: 10.1007/978-1-4939-8778-8_20.
With ever-increasing genomic information combined with modern tools for genome modification, we are entering a new era of plant biotechnology. One major tool used for genome modification is the zinc finger nuclease (ZFN). Here, we discuss how ZFNs have proven useful in many genome modification applications. In order to remove the function of a gene or genes, targeted mutagenesis using ZFNs has been readily demonstrated creating numerous gene knockouts, and gene deletion has been demonstrated with removal of gene segments both native and transgenic up to 9 Mb. Applications for gain of function have also been demonstrated. Precision gene editing using ZFNs has resulted in the development of herbicide tolerance, and numerous forms of targeted gene addition have been exhibited. In addition to genome modification, this chapter also describes the use of zinc finger protein transcription factors (ZFP-TFs) for gene regulation in order to provide useful modification of gene expression resulting in altered phenotypes.
随着基因组信息不断增加并与现代基因组编辑工具相结合，我们正步入植物生物技术的新时代。用于基因组编辑的一种主要工具是锌指核酸酶（ZFN）。在此，我们将探讨ZFN在许多基因组编辑应用中已被证明是如何发挥作用的。为了去除一个或多个基因的功能，利用ZFN进行的靶向诱变已很容易地实现了大量基因敲除，并且通过去除长达9 Mb的天然和转基因基因片段已证明了基因删除。功能获得的应用也已得到证实。使用ZFN进行的精确基因编辑已带来了除草剂耐受性的发展，并且已展示出多种形式的靶向基因添加。除了基因组编辑外，本章还描述了使用锌指蛋白转录因子（ZFP-TF）进行基因调控，以便对基因表达进行有效修饰从而导致表型改变。
