Department of Plant Sciences, University of Tennessee, Knoxville, TN, 37996, USA.
Department of Food Science, University of Tennessee, Knoxville, TN, 37996, USA; Department of Mechanical, Aerospace, and Biomedical Engineering, University of Tennessee, Knoxville, TN, 37996, USA.
Plant Sci. 2018 Aug;273:42-49. doi: 10.1016/j.plantsci.2018.02.025. Epub 2018 Mar 3.
Genome editing is a powerful suite of technologies utilized in basic and applied plant research. Both nuclear and plastid genomes have been genetically engineered to alter traits in plants. While the most frequent molecular outcome of gene editing has been knockouts resulting in a simple deletion of an endogenous protein of interest from the host's proteome, new genes have been added to plant genomes and, in several instances, the sequence of endogenous genes have been targeted for a few coding changes. Targeted plant characteristics for genome editing range from single gene targets for agronomic input traits to metabolic pathways to endow novel plant function. In this paper, we review the fundamental approaches to editing nuclear and plastid genomes in plants with an emphasis on those utilizing synthetic biology. The differences between the eukaryotic-type nuclear genome and the prokaryotic-type plastid genome (plastome) in plants has profound consequences in the approaches employed to transform, edit, select transformants, and indeed, nearly all aspects of genetic engineering procedures. Thus, we will discuss the two genomes targeted for editing in plants, the toolbox used to make edits, along with strategies for future editing approaches to transform crop production and sustainability. While CRISPR/Cas9 is the current method of choice in editing nuclear genomes, the plastome is typically edited using homologous recombination approaches. A particularly promising synthetic biology approach is to replace the endogenous plastome with a 'synplastome' that is computationally designed, and synthesized and assembled in the lab, then installed into chloroplasts. The editing strategies, transformation methods, characteristics of the novel plant also affect how the genetically engineered plant may be governed and regulated. Each of these components and final products of gene editing affect the future of biotechnology and farming.
基因组编辑是一套在基础和应用植物研究中广泛应用的强大技术。核基因组和质体基因组都经过遗传工程改造，以改变植物的特性。虽然基因编辑最常见的分子结果是敲除，导致内源目的蛋白从宿主蛋白质组中简单缺失，但新的基因已被添加到植物基因组中，并且在几个实例中，内源基因的序列已被靶向进行少数编码改变。基因组编辑的靶向植物特征范围从单一基因目标的农艺投入性状到代谢途径，以赋予新的植物功能。在本文中，我们综述了利用合成生物学编辑植物核基因组和质体基因组的基本方法，重点介绍那些方法。植物真核型核基因组和原核型质体基因组（质体）之间的差异对转化、编辑、选择转化体的方法以及实际上几乎所有遗传工程程序的方面都有深远的影响。因此，我们将讨论植物中编辑的两个基因组、用于进行编辑的工具箱以及用于改造作物生产和可持续性的未来编辑方法的策略。虽然 CRISPR/Cas9 是编辑核基因组的当前首选方法，但质体通常使用同源重组方法进行编辑。一种特别有前途的合成生物学方法是用经过计算设计、在实验室中合成和组装的“合成质体”取代内源质体，然后将其安装到叶绿体中。编辑策略、转化方法、新型植物的特征也会影响基因工程植物的管理和监管方式。基因编辑的这些组件和最终产物中的每一个都影响着生物技术和农业的未来。
