Lycopene Is Enriched in Tomato Fruit by CRISPR/Cas9-Mediated Multiplex Genome Editing

Li, Xindi; Wang, Yanning; Chen, Sha; Tian, Huiqin; Fu, Daqi; Zhu, Beiwei; Luo, Yunbo; Zhu, Hongliang

doi:10.3389/fpls.2018.00559

Cited by 261 publications

(110 citation statements)

References 60 publications

(89 reference statements)

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“…Another strategy is to suppress competing metabolic pathways. Using a multiplex pYLCRISPR/Cas9 system that targeted five key genes, the γ‐aminobutyric acid (GABA) shunt was converted in tomato (Li et al , ), and lycopene was enriched in tomato fruit by knocking down five genes associated with the carotenoid metabolic pathway (Li et al , ).…”

Section: Discussionmentioning

confidence: 99%

Multiplex CRISPR/Cas9‐mediated metabolic engineering increases soya bean isoflavone content and resistance to soya bean mosaic virus

Zhang

Wang

et al. 2019

Plant Biotechnology Journal

173

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and (Tel 86-25-84396410; fax 86-25-84396410; email dyyu@njau.edu.cn) † These authors contribute equally to this work. SummaryIsoflavonoids, which include a variety of secondary metabolites, are derived from the phenylpropanoid pathway and are distributed predominantly in leguminous plants. These compounds play a critical role in plant-environment interactions and are beneficial to human health. Isoflavone synthase (IFS) is a key enzyme in isoflavonoid synthesis and shares a common substrate with flavanone-3-hydroxylase (F3H) and flavone synthase II (FNS II). In this study, CRISPR/Cas9-mediated multiplex gene-editing technology was employed to simultaneously target GmF3H1, GmF3H2 and GmFNSII-1 in soya bean hairy roots and plants. Various mutation types and frequencies were observed in hairy roots. Higher mutation efficiencies were found in the T 0 transgenic plants, with a triple gene mutation efficiency of 44.44%, and these results of targeted mutagenesis were stably inherited in the progeny. Metabolomic analysis of T 0 triplemutants leaves revealed significant improvement in isoflavone content. Compared with the wild type, the T 3 generation homozygous triple mutants had approximately twice the leaf isoflavone content, and the soya bean mosaic virus (SMV) coat protein content was significantly reduced by one-third after infection with strain SC7, suggesting that increased isoflavone content enhanced the leaf resistance to SMV. The isoflavone content in the seeds of T 2 triple mutants was also significantly increased. This study provides not only materials for the improvement of soya bean isoflavone content and resistance to SMV but also a simple system to generate multiplex mutations in soya bean, which may be beneficial for further breeding and metabolic engineering. 1384

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Section: Discussionmentioning

confidence: 99%

Multiplex CRISPR/Cas9‐mediated metabolic engineering increases soya bean isoflavone content and resistance to soya bean mosaic virus

Zhang

Wang

et al. 2019

Plant Biotechnology Journal

173

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“…In yet another study in Camelina , the content of very long fatty acids was reduced by knocking out the FAE1 gene (Ozseyhan et al ). An enrichment of the nutritionally highly valuable compound lycopene was achieved in tomato fruits via multiplex site‐directed mutagenesis of several genes involved in the carotenoid biosynthesis pathway (Li et al ).…”

Section: Applications Of Targeted Genome Modification By Nhejmentioning

confidence: 99%

The CRISPR/Cas revolution continues: From efficient gene editing for crop breeding to plant synthetic biology

Kumlehn

Pietralla

Hensel

et al. 2018

JIPB

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Since the discovery that nucleases of the bacterial CRISPR (clustered regularly interspaced palindromic repeat)‐associated (Cas) system can be used as easily programmable tools for genome engineering, their application massively transformed different areas of plant biology. In this review, we assess the current state of their use for crop breeding to incorporate attractive new agronomical traits into specific cultivars of various crop plants. This can be achieved by the use of Cas9/12 nucleases for double‐strand break induction, resulting in mutations by non‐homologous recombination. Strategies for performing such experiments − from the design of guide RNA to the use of different transformation technologies − are evaluated. Furthermore, we sum up recent developments regarding the use of nuclease‐deficient Cas9/12 proteins, as DNA‐binding moieties for targeting different kinds of enzyme activities to specific sites within the genome. Progress in base deamination, transcriptional induction and transcriptional repression, as well as in imaging in plants, is also discussed. As different Cas9/12 enzymes are at hand, the simultaneous application of various enzyme activities, to multiple genomic sites, is now in reach to redirect plant metabolism in a multifunctional manner and pave the way for a new level of plant synthetic biology.

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“…Using CRISPR-Cas genome editing, the nutritional profiles of many crops have been recently 172 demonstrated. For example, in tomato, knocking down genes in the carotenoid metabolic pathway led 173 to a 5-fold increase in lycopene (Li et al, 2018); and in rice, generating mutations in the starch 174 branching enzyme (Berrens et al 2017) genes increased amylose content by up to 25% and resistant 175 starch to 9.8% (Sun et al, 2017). One of the most significant impacts of CRISPR-Cas genome editing 176 is the potential improvement of a key trait in a commercially released cultivar within 6 months.…”

Section: Application Of Biotechnology To Produce Zero-toxin Vetch 158mentioning

confidence: 99%

Common Vetch: a drought tolerant, high protein neglected leguminous crop with potential as a sustainable food source

Nguyen

Riley

Nagel

et al. 2020

Preprint

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13• Global demand for protein is predicted to increase by 50% by 2050. To meet the increasing demand 14 whilst ensuring sustainability, protein sources that generate low-greenhouse gas emissions are required 15 and protein-rich legume seeds have the potential to make a significant contribution. Legumes like 16 common vetch (Vicia sativa) that grow in marginal cropping zones and are drought tolerant and 17 resilient to changeable annual weather patterns, will be in high demand as the climate changes. In 18 common vetch, the inability to eliminate the γ-glutamyl-β-cyano-alanine (GBCA) toxin present in the 19 seed has hindered its utility as a human and animal food for many decades, leaving this highly resilient 20 species an "orphan" legume. However, the availability of the vetch genome and transcriptome data 21 together with the application of CRISPR-Cas genome editing technologies lay the foundations to 22 eliminate the GBCA toxin constraint. In the near future, we anticipate that a zero-toxin vetch variety 23 will become a significant contributor to global protein demand. 24

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Lycopene Is Enriched in Tomato Fruit by CRISPR/Cas9-Mediated Multiplex Genome Editing

Cited by 261 publications

References 60 publications

Multiplex CRISPR/Cas9‐mediated metabolic engineering increases soya bean isoflavone content and resistance to soya bean mosaic virus

Multiplex CRISPR/Cas9‐mediated metabolic engineering increases soya bean isoflavone content and resistance to soya bean mosaic virus

The CRISPR/Cas revolution continues: From efficient gene editing for crop breeding to plant synthetic biology

Common Vetch: a drought tolerant, high protein neglected leguminous crop with potential as a sustainable food source

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