Abstract：
CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated) technology is a versatile genome editing tool that has been used to improve agriculturally important plant traits. Due to its precision, CRISPR/Cas9 is more effective than either conventional plant breeding methods or standard genetic engineering approaches for the rapid development of new varieties resilient to climate change. In addition to knowledge in tissue culture-based plant transformation, effective gene-specific single guide RNA (sgRNA) design, prediction of its off-target effect and utilization of vectors, promoters, Cas proteins and terminators is required for CRISPR/Cas9. Various bioinformatics tools are available for the best sgRNA design and screening of the off-targets. Various tools are used in the delivery of CRISPR/Cas components into cells and the genome. Moreover, some recent studies proved the simultaneous silencing of different paralogs in the same family or several genes working in the same pathway by using multiple-target sgRNA designs. This review summarizes the type of promoters, Cas proteins, recognition sequences, and terminators available for the development of knock-out and overexpression plant lines. It also provides a general guideline for the development of genome-edited plants from the design of sgRNAs to the selection of non-transgenic genome-edited T2 generation.
摘要：
CRISPR（成簇的规则间隔的短回文重复）/Cas（CRISPR相关）技术是一种通用的基因组编辑工具，已用于改善农业上重要的植物性状。由于其精确性,CRISPR/Cas9比传统植物育种方法或标准基因工程方法更有效地快速开发适应气候变化的新品种。除了在组织培养为基础的植物转化的知识，有效的基因特异性单指导RNA（sgRNA）设计，预测其离靶效应和矢量利用率，promotors,CRISPR/Cas9需要Cas蛋白和终止子。各种生物信息学工具可用于最佳sgRNA设计和脱靶筛选。各种工具用于将CRISPR/Cas组分递送到细胞和基因组中。此外，最近的一些研究证明，通过使用多靶sgRNA设计，同一家族中的不同旁系同源物或在同一途径中工作的几个基因同时沉默。这篇综述总结了发起人的类型，Cas蛋白质，识别序列,和可用于开发敲除和过表达植物品系的终止子。它还提供了从sgRNA的设计到非转基因基因组编辑的T2代的选择的基因组编辑的植物的开发的一般指南。
