Abstract
The emergence of genome-editing technology has allowed manipulation of DNA sequences in genomes to precisely remove or replace specific sequences in organisms resulting in targeted mutations. In plants, genome editing is an attractive method to alter gene functions to generate improved crop varieties. Genome editing is thought to be simple to use and has a lower risk of off-target effects compared to classical mutation breeding. Furthermore, genome-editing technology tools can also be applied directly to crops that contain complex genomes and/or are not easily bred using traditional methods. Currently, highly versatile genome-editing tools for precise and predictable editing of almost any locus in the plant genome make it possible to extend the range of application, including functional genomics research and molecular crop breeding. Vegetables are essential nutrient sources for humans and provide vitamins, minerals, and fiber to diets, thereby contributing to human health. In this review, we provide an overview of the brief history of genome-editing technologies and the components of genome-editing tool boxes, and illustrate basic modes of operation in representative systems. We describe the current and potential practical application of genome editing for the development of improved nutritious vegetables and present several case studies demonstrating the potential of the technology. Finally, we highlight future directions and challenges in applying genome-editing systems to vegetable crops for research and product development.
Highlights
There has been growing interest in the beneficial effect of consuming vegetables because of the broad range of nutritional compounds, such as vitamins, minerals, antioxidants, dietary fiber, and a plethora of phytochemical compounds present in this crop group (Septembre-Malaterre et al, 2018)
Mutagenic chemicals were preferred for mutagenesis because of easier handling and higher mutation frequency compared to radiological methods
The most commonly used delivery methods for DNA constructs carrying the clustered regulatory interspaced short palindromic repeats (CRISPR)-cas9 components are biolistic bombardment, Agrobacterium-mediated delivery to explants or plants, and direct transfer of the constructs to protoplasts with polyethylene glycol (PEG), which have been successful for generating CRISPRCas-mediated genome editing in various plant species with varying efficiencies of transformation (Borrelli et al, 2018)
Summary
There has been growing interest in the beneficial effect of consuming vegetables because of the broad range of nutritional compounds, such as vitamins, minerals, antioxidants, dietary fiber, and a plethora of phytochemical compounds present in this crop group (Septembre-Malaterre et al, 2018). Advanced molecular biological methods using SSNs, such as zinc-finger nucleases (ZFNs) and transcription activatorlike effector nucleases (TALENs), and CRISPR-Cas system (Kim et al, 1996; Christian et al, 2010; Jinek et al, 2012; Chen and Gao, 2014; Gao, 2014), have made it possible for plant researchers to conduct targeted gene/genome engineering precisely and efficiently These techniques generate double-strand breaks (DSBs) at specific DNA sites and, via the endogenous DNA repair system, induce insertions or deletions of nucleotides by non-homologous end joining (NHEJ), or cause gene replacements by homologous recombination (HR) thereby resulting in loss-of-function or gain-of-function of target genes (Symington and Gautier, 2011; Figure 1). It allows producing specific mutations effectively in a selected genetic background
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