Construct design for CRISPR/Cas-based genome editing in plants.

Authors :: Hassan MM
Zhang Y
Yuan G
De K
Chen JG
Muchero W
Tuskan GA
Qi Y
Yang X
Source :: Trends in plant science [Trends Plant Sci] 2021 Nov; Vol. 26 (11), pp. 1133-1152. Date of Electronic Publication: 2021 Jul 31.
Publication Year :: 2021
Abstract: 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.<br />Competing Interests: Declaration of interests Y.Q. is a consultant for Inari Agriculture and CTC Genomics. The remaining authors declare no conflict of interests.<br /> (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)