Joo Kathleen A, Muszynski Michael G, Kantar Michael B, Wang Ming-Li, He Xiaoling, Del Valle Echevarria Angel R
Department of Tropical Plant and Soil Sciences, University of Hawaii at Manoa, Honolulu, HI, United States.
Hawaii Agriculture Research Center, Waipahu, HI, United States.
Front Genet. 2021 Nov 12;12:786140. doi: 10.3389/fgene.2021.786140. eCollection 2021.
Adopting modern gene-editing technologies for trait improvement in agriculture requires important workflow developments, yet these developments are not often discussed. Using tropical crop systems as a case study, we describe a workflow broken down into discrete processes with specific steps and decision points that allow for the practical application of the CRISPR-Cas gene editing platform in a crop of interest. While we present the steps of developing genome-edited plants as sequential, in practice parts can be done in parallel, which are discussed in this perspective. The main processes include 1) understanding the genetic basis of the trait along with having the crop's genome sequence, 2) testing and optimization of the editing reagents, development of efficient 3) tissue culture and 4) transformation methods, and 5) screening methods to identify edited events with commercial potential. Our goal in this perspective is to help any lab that wishes to implement this powerful, easy-to-use tool in their pipeline, thus aiming to democratize the technology.
将现代基因编辑技术应用于农业性状改良需要重要的工作流程改进,但这些改进往往未被讨论。以热带作物系统为例,我们描述了一个细分为离散过程的工作流程,这些过程具有特定步骤和决策点,可实现CRISPR-Cas基因编辑平台在目标作物中的实际应用。虽然我们将基因组编辑植物的开发步骤按顺序呈现,但实际上部分步骤可以并行进行,本文将对此进行讨论。主要过程包括:1)了解性状的遗传基础并掌握作物的基因组序列;2)测试和优化编辑试剂,开发高效的3)组织培养和4)转化方法,以及5)筛选方法以识别具有商业潜力的编辑事件。我们撰写本文的目的是帮助任何希望在其流程中应用这一强大且易于使用工具的实验室,从而致力于使该技术普及。