Construct design for CRISPR/Cas-based genome editing in plants
Md Mahmudul Hassan, Yingxiao Zhang, Guoliang Yuan, Juntal De, Jin-Gui Chen, Wellington Muchero, Gerald A. Tuskan, Yiping Qi and Xiaohan Yang
31 July 2021, Trends in Plant Sciences 26(11): 1133-1152; https://doi.org/10.1016/j.tplants.2021.06.015
CRISPR construct design is a key step in the practice of genome editing, which includes identification of appropriate Cas proteins, design and selection of guide RNAs (gRNAs), and selection of regulatory elements to express gRNAs and Cas proteins. Here, we review the choices of CRISPR-based genome editors suited for different needs in plant genome editing applications. We consider the technical aspects of gRNA design and the associated computational tools. We also discuss strategies for the design of multiplex CRISPR constructs for high-throughput manipulation of complex biological processes or polygenic traits. We provide recommendations for different elements of CRISPR constructs and discuss the remaining challenges of CRISPR construct optimization in plant genome editing.