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Review
. 2021 Apr 3;22(7):3743.
doi: 10.3390/ijms22073743.

Application of Upstream Open Reading Frames (uORFs) Editing for the Development of Stress-Tolerant Crops

Taeyoung Um  1 , Taehyeon Park  2 , Jae Sung Shim  3 , Youn Shic Kim  1 , Gang-Seob Lee  4 , Ik-Young Choi  5 , Ju-Kon Kim  2 , Jun Sung Seo  2 , Soo Chul Park  4
Affiliations
Review

Application of Upstream Open Reading Frames (uORFs) Editing for the Development of Stress-Tolerant Crops

Taeyoung Um et al. Int J Mol Sci. .

Abstract

Global population growth and climate change are posing increasing challenges to the production of a stable crop supply using current agricultural practices. The generation of genetically modified (GM) crops has contributed to improving crop stress tolerance and productivity; however, many regulations are still in place that limit their commercialization. Recently, alternative biotechnology-based strategies, such as gene-edited (GE) crops, have been in the spotlight. Gene-editing technology, based on the clustered regularly interspaced short palindromic repeats (CRISPR) platform, has emerged as a revolutionary tool for targeted gene mutation, and has received attention as a game changer in the global biotechnology market. Here, we briefly introduce the concept of upstream open reading frames (uORFs) editing, which allows for control of the translation of downstream ORFs, and outline the potential for enhancing target gene expression by mutating uORFs. We discuss the current status of developing stress-tolerant crops, and discuss uORF targets associated with salt stress-responsive genes in rice that have already been verified by transgenic research. Finally, we overview the strategy for developing GE crops using uORF editing via the CRISPR-Cas9 system. A case is therefore made that the mutation of uORFs represents an efficient method for developing GE crops and an expansion of the scope of application of genome editing technology.

Keywords: CRISPR; enhancing the gene expression; gene editing; stress-resistant crops; upstream open reading frames (uORFs).

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Conflict of interest statement

All authors declare that there is no conflict of interest.

Figures

Figure 1
Figure 1
Overview of CRISPR-Cas9-mediated regulation of the repression of translation, as discussed in this review. (A) The mRNA (black rectangle) with uORF (gray rectangle) induces ribosome (yellow ovals) stalling in the uORF, which represses translation of the main ORF (mORF). Polypeptide: gray circle. (B) Mutation of the start codon region (green or red rectangle) in uORF using CRISPR-Cas9 inhibits ribosome stalling, leading to induced translation of mORF.
Figure 2
Figure 2
Regulation of uORF-mediated translation and types of uORF. (A) The various mechanism of translational repression. (B) Types of uORFs. uORFs are classified multiple type and type 1–3. Blue, gray and black rectangles indicate regions of the mRNA, yellow ovals indicate the ribosome complex, and orange circles indicate the exosome.
Figure 3
Figure 3
Mechanism of avoiding uORF-mediated repression of gene expression. Alternative splicing (A) and alternative transcription start site (TSS) (B) to exclude a uORF from mRNA.
Figure 4
Figure 4
Overview of the process of gene editing of uORFs with the CRISPR system for the development of GE crops.
See this image and copyright information in PMC

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