Abstract
The advent of precise genome-editing tools has revolutionized the way we create new plant varieties. Three groups of tools are now available, classified according to their mechanism of action: Programmable sequence-specific nucleases, base-editing enzymes, and oligonucleotides. The corresponding techniques not only lead to different outcomes, but also have implications for the public acceptance and regulatory approval of genome-edited plants. Despite the high efficiency and precision of the tools, there are still major bottlenecks in the generation of new and improved varieties, including the efficient delivery of the genome-editing reagents, the selection of desired events, and the regeneration of intact plants. In this review, we evaluate current delivery and regeneration methods, discuss their suitability for important crop species, and consider the practical aspects of applying the different genome-editing techniques in agriculture.
Highlights
The precise modification of pre-selected sequences in the plant genome is the holy grail of plant breeding because it allows the rapid introduction of genetic diversity and accelerates the generation of improved varieties, especially in polyploid crops, which otherwise need to undergo lengthy programs of crossing and screening
There are three major genome-editing techniques categorized by mechanism of action, and by far the most commonly used in plants is the targeted generation of DNA double strand breaks (DSBs) using programmable sequence-specific nucleases followed by DNA repair via one of two major endogenous pathways [1,2]
Mutagenic DNA oligonucleotides 20–200 nucleotides in length have been delivered into plant cells to introduce point mutations in target genes, an approach known as oligonucleotide-directed mutagenesis (ODM)
Summary
The precise modification of pre-selected sequences in the plant genome is the holy grail of plant breeding because it allows the rapid introduction of genetic diversity and accelerates the generation of improved varieties, especially in polyploid crops, which otherwise need to undergo lengthy programs of crossing and screening. The development and application of genome-editing tools in recent years has, revolutionized basic research in plant biology and the generation of new plant varieties. The molecular basis of genome editing in plants has been comprehensively discussed in the literature, the practical aspects have received comparatively little attention. In this article, we emphasize some of the technical and practical aspects of genome-editing technologies for crop improvement
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