Abstract

Abstract Improving crops through plant breeding, an important approach for sustainable agriculture, has been utilized to increase the yield and quality of foods and other biomaterials for human use. Crops, including cereals, vegetables, ornamental flowers, fruits, and trees, have long been cultivated to produce high-quality products for human consumption. Conventional breeding technologies, such as natural cross-hybridization, mutation induction through physical or chemical mutagenesis, and modern transgenic tools are often used to enhance crop production. However, these breeding methods are sometimes laborious and complicated, especially when attempting to improve desired traits without inducing pleiotropic effects. Recently, targeted genome editing (TGE) technology using engineered nucleases, including meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR) nucleases, has been used to improve the traits of economically important plants. TGE has emerged as a novel plant-breeding tool that represents an alternative approach to classical breeding, but with higher mutagenic efficiency. Here, we briefly describe the basic principles of TGE and the types of engineered nucleases utilized, along with their advantages and disadvantages. We also discuss their potential use to improve the traits of horticultural crops through genome engineering.

Highlights

  • Plant breeding is a classical agricultural activity that has been practiced for thousands of years

  • targeted genome editing (TGE) mainly relies on four types of engineered nucleases (EN), including homing endonucleases or meganucleases (EMNs), zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspersed short palindromic repeats (CRISPR)/Cas9 reagents (Gaj et al, 2013)

  • When a Cas9 nuclease binds to its target site on a DNA strand, it undergoes a structural change, which places its domains on the opposite strands to create a double-stranded breaks (DSBs) at a site three nucleotides before protospacer adjacent motif (PAM) (Fig. 2D) in the target DNA

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Summary

Introduction

Plant breeding is a classical agricultural activity that has been practiced for thousands of years. TGE mainly relies on four types of ENs, including homing endonucleases or meganucleases (EMNs), zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspersed short palindromic repeats (CRISPR)/Cas reagents (Gaj et al, 2013) These ENs contain sequence-specific DNA-binding domains, which cleave nucleotides in the target genes in a sequence-specific manner, thereby generating double-stranded breaks (DSBs) (Gaj et al, 2013; Carroll, 2014). When a Cas nuclease binds to its target site on a DNA strand, it undergoes a structural change, which places its domains on the opposite strands to create a DSB at a site three nucleotides before PAM (Fig. 2D) in the target DNA These DSBs can subsequently be repaired by the HDR or NHEJ mechanism, resulting in InDel mutations at the target sites. RNAi technology has many limitations, such as the reduced frequency of gene silencing, the induction of unwanted pathways, and high costs

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