Agrobacterium -mediated delivery of CRISPR/Cas reagents for genome editing in plants enters an era of ternary vector sys
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robacterium-mediated delivery of CRISPR/Cas reagents for genome editing in plants enters an era of ternary vector systems 1
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Yu Zhang , Qiang Zhang & Qi-Jun Chen 1
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, 2
Beijing 100193, China; Center for Crop Functional Genomics and Molecular Breeding, China Agricultural University, Beijing 100193, China Received March 6, 2020; accepted March 19, 2020; published online March 27, 2020
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. genome editing, CRISPR/Cas, ternary vector system, delivery, Agrobacterium-mediated transformation Citation:
Zhang, Y., Zhang, Q., and Chen, Q.J. (2020). Agrobacterium-mediated delivery of CRISPR/Cas reagents for genome editing in plants enters an era of ternary vector systems. Sci China Life Sci 63, https://doi.org/10.1007/s11427-020-1685-9
Introduction CRISPR/Cas-based tools have revolutionized life sciences, providing unprecedented opportunities for crop breeding and functional genomics research (Meng et al., 2018; Xue et al., 2018; Chen et al., 2019; Ji et al., 2019; Li et al., 2019; Liu et al., 2019). However, lack of appropriate methods to deliver genome-editing reagents (i.e., constructs encoding the nuclease and target-specific RNAs) is the major barrier to CRISPR/Cas-mediated gene editing in a variety of plants (Ran et al., 2017). Agrobacterium-mediated genetic transformation (AMGT) is the preferred method of CRISPR/Cas
reagent delivery; however, the process typically remains confined to limited species and genotypes, often non-elite cultivars with poor agronomic characteristics and low efficiencies of delivery (Altpeter et al., 2016; Lowe et al., 2016). Fortunately, the recent use of ternary vector systems coupled with morphogenic or developmental regulator (MR or DR) genes has greatly improved AMGT methods (Lowe et al., 2016; Anand et al., 2018; Che et al., 2018; Lowe et al., 2018; Zhang et al., 2019).
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