Methods and Techniques to Select Efficient Guides for CRISPR-Mediated Genome Editing in Plants

Abstract

CRISPR technology is revolutionizing genomic engineering by enabling scientists to precisely modify plant DNA, thus representing a powerful tool for plant breeding.This chapter provides a summary of the approaches and constraints of CRISPR-mediated genome editing in plants, with a focus on the critical prerequisite of efficient CRISPR reagents for successful gene editing in plants.While computational tools have tremendously improved our ability to design specific guides, their limitations make guide effectiveness prediction unreliable, especially for plants. Therefore, it is strongly recommended to validate CRISPR reagents before investing time and resources in the challenging process of plant transformation.A number of in vitro and in planta assays coupled with analytical methods have been proposed to assess the editing performances. Each approach has its own strengths and weaknesses, so the choice of the most suitable system depends on the specific plant species and the type and depth of the genotypic data required.In many cases, the hairy root assay can provide a good compromise between rapidity, reliability and cost-effectiveness for assessing editing performance in numerous plant species.