State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
Center for Crop Functional Genomics and Molecular Breeding, China Agricultural University, Beijing, 100193, China.
Sci China Life Sci. 2020 Oct;63(10):1491-1498. doi: 10.1007/s11427-020-1685-9. Epub 2020 Mar 27.
Lack of appropriate methods for delivery of genome-editing reagents is a major barrier to CRISPR/Cas-mediated genome editing in plants. Agrobacterium-mediated genetic transformation (AMGT) is the preferred method of CRISPR/Cas reagent delivery, and researchers have recently made great improvements to this process. In this article, we review the development of AMGT and AMGT-based delivery of CRISPR/Cas reagents. We give an overview of the development of AMGT vectors including binary vector, superbinary vector, dual binary vector, and ternary vector systems. We also review the progress in Agrobacterium genomics and Agrobacterium genetic engineering for optimal strains. We focus in particular on the ternary vector system and the resources we developed. In summary, it is our opinion that Agrobacterium-mediated CRISPR/Cas genome editing in plants is entering an era of ternary vector systems, which are often integrated with morphogenic regulators. The new vectors described in this article are available from Addgene and/or MolecularCloud for sharing with academic investigators for noncommercial research.
缺乏适当的基因组编辑试剂传递方法是 CRISPR/Cas 介导的植物基因组编辑的主要障碍。农杆菌介导的遗传转化 (AMGT) 是 CRISPR/Cas 试剂传递的首选方法，研究人员最近对此过程进行了重大改进。在本文中，我们回顾了 AMGT 的发展以及基于 AMGT 的 CRISPR/Cas 试剂传递。我们概述了 AMGT 载体的发展，包括二元载体、超级二元载体、双二元载体和三元载体系统。我们还回顾了用于优化菌株的农杆菌基因组学和农杆菌遗传工程的进展。我们特别关注三元载体系统和我们开发的资源。总之，我们认为农杆菌介导的植物 CRISPR/Cas 基因组编辑正在进入三元载体系统的时代，这些系统通常与形态发生调节剂整合在一起。本文中描述的新载体可从 Addgene 和/或 MolecularCloud 获得，以供学术研究人员用于非商业性研究的共享。
