期刊
TRENDS IN PLANT SCIENCE
卷 26, 期 11, 页码 1133-1152出版社
CELL PRESS
DOI: 10.1016/j.tplants.2021.06.015
关键词
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资金
- Center for Bioenergy Innovation (CBI), a U.S. Department of Energy (DOE) Research Centre
- Biological and Environmental Research (BER) program, the Laboratory Directed Research and Development (LDRD) program of Oak Ridge National Laboratory
- U.S. DOE BER Genomic Science Program
- Secure Ecosystem Engineering and Design (SEED) Scientific Focus Areas
- U.S. Department of Energy
- National Science Foundation Plant Genome Research Program [IOS-1758745, IOS-2029889]
- U.S. Department of Agriculture Biotechnology Risk Assessment Grant Program competitive grants [2018-33522-28789, 2020-33522-32274]
- Emergency Citrus Disease Research and Extension Program [2020-70029-33161, 2021-67013-34554]
- Foundation for Food and Agriculture Research grant [593603]
- UT-Battelle, LLC [DE-AC05-00OR22725]
CRISPR construct design is essential for genome editing, involving selection of Cas proteins, gRNAs, and regulatory elements. This review covers different options for CRISPR-based genome editors in plant editing, gRNA design, multiplex construct strategies, and challenges in construct optimization.
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 highthroughput 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.
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