CRISPR–Cas9-mediated genome editing in apple and grapevine

Osakabe, Yuriko; Liang, Zhenchang; Ren, Chong; Nishitani, Chikako; Osakabe, Keishi; Wada, Masato; Komori, Sadao; Malnoy, Mickaël; Velasco, Riccardo; Poli, Michele; Jung, Minhee; Koo, Ok Jae; Viola, R.; Kanchiswamy, Chidananda Nagamangala

doi:10.1038/s41596-018-0067-9

Cited by 161 publications

(124 citation statements)

References 43 publications

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“…The application of these new technologies may be particularly useful in grapevine since they produce minimal and precise modifications in selected genotypes of interest, such as elite cultivars sought-after by the wine market, without altering the genetic background as happens in traditional breeding. CRISPR/Cas9 technology has been successfully applied to generate edited grapevine plants (Osakabe et al, 2018). Two different delivery systems have been used: one based on the stable integration of the genetic components in the genome by means of A. tumefaciens gene transfer and the second one based on the direct delivery of purified Cas9 protein and gRNAs (Figure 4).…”

Section: State-of-the Art Of the Nbtsmentioning

confidence: 99%

The state-of-the-art of grapevine biotechnology and new breeding technologies (NBTS)

et al. 2019

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Context of the review:The manipulation of the genetic basis controlling grapevine adaptation and phenotypic plasticity can be performed either by classical genetics or biotechnologies. In the last 15 years, considerable knowledge has accumulated about the grapevine genome as well as the mechanisms involved in the interaction of the vine with the environment, pests and diseases. Despite the difficulties associated with genetic mapping in this species (allele diversity, chimerism, long generation intervals…), several major QTLs (quantitative trait loci) controlling important vegetative or reproductive traits have been identified. Considering the huge genotypic and phenotypic diversities existing in Vitis, breeding offers a substantial range of options to improve the performances of cultivars. However, even if marker-assisted selection was largely developed to shorten breeding programs, the selection of improved cultivars, whether for agronomic traits or disease tolerances, is still long and uncertain. Moreover, breeding by crossing does not preserve cultivar genetic background, when the wine industry and market are still based on varietal wines. Significance of the review: In grapevine, pioneering biotechnologies were set up in the 1960s to propagate and/or clean the material from micro-organisms. In the 1990s, the basis of genetic engineering was primary established through biolistic or Agrobacterium with several derived technologies refined in the last 10 years. The latest advance is represented by a group of technologies based on genome editing which allows a much more precise modification of the genome. These technologies, so-called NBTs (new breeding technologies), which theoretically do not deconstruct the phenotype of existing cultivars, could be potentially better accepted by the wine industry and consumers than previous GMO (genetically modified organism) approaches. This paper reviews the current state-ofthe-art of the biotechnologies available for grapevine genome manipulation and future prospects for genetic improvement.

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Section: State-of-the Art Of the Nbtsmentioning

confidence: 99%

The state-of-the-art of grapevine biotechnology and new breeding technologies (NBTS)

et al. 2019

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“…However, no plants were regenerated. Osakabe et al [91] also developed a stepwise protocol for the design and transfer of CRISPR-Cas9 components with high accuracy and efficiency in apple and grapevine protoplasts. The complete protocol employing the direct delivery of CRISPR-Cas9 RNPs takes as little as 2-3 weeks, with the advantage of generating exogenous DNA-free plants, whereas the plasmid-mediated procedure takes more than 3 months to regenerate plants and study the mutations.…”

Section: Crispr/cas9 Genome Editing For Generation Of Non-transgenic mentioning

confidence: 99%

Development of Improved Fruit, Vegetable, and Ornamental Crops Using the CRISPR/Cas9 Genome Editing Technique

Corte

Mahmoud

Moraes

et al. 2019

Plants

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Horticultural crops, including fruit, vegetable, and ornamental plants are an important component of the agriculture production systems and play an important role in sustaining human life. With a steady growth in the world's population and the consequent need for more food, sustainable and increased fruit and vegetable crop production is a major challenge to guarantee future food security. Although conventional breeding techniques have significantly contributed to the development of important varieties, new approaches are required to further improve horticultural crop production. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) has emerged as a valuable genome-editing tool able to change DNA sequences at precisely chosen loci. The CRISPR/Cas9 system was developed based on the bacterial adaptive immune system and comprises of an endonuclease guided by one or more single-guide RNAs to generate double-strand breaks. These breaks can then be repaired by the natural cellular repair mechanisms, during which genetic mutations are introduced. In a short time, the CRISPR/Cas9 system has become a popular genome-editing technique, with numerous examples of gene mutation and transcriptional regulation control in both model and crop plants. In this review, various aspects of the CRISPR/Cas9 system are explored, including a general presentation of the function of the CRISPR/Cas9 system in bacteria and its practical application as a biotechnological tool for editing plant genomes, particularly in horticultural crops.

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“…Among them, the CRISPR/Cas9 system is predominantly used for TGE due to its simplicity, high efficiency and versatility [8]. In most cases, the Cas9 encoding gene is driven by constitutive promoters, such as CaMV 35S and ubiquitin promoters [9,10,11,12,13]. However, constitutive expression of Cas9 might be harmful to cells, or increase the risk of off-target effect [14,15].…”

Section: Introductionmentioning

confidence: 99%

Targeted genome editing inNicotiana tabacumusing inducible CRISPR/Cas9 system

Ren

Wang

et al. 2020

Preprint

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Targeted genome editing has been achieved in multiple plant species using the clustered 10 regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (CRISPR/Cas9) 11 system, in which the Cas9 gene is usually driven by constitutive promoters. However, constitutive 12 expression of Cas9 is not necessary and can be harmful to plant development. In this study, we 13 developed an estrogen-inducible CRISPR/Cas9 system by taking advantage of the chimeric 14 transcription activator XVE and tested the efficacy of this inducible system in Nicotiana tabacum by 15 targeting the phytoene desaturase (NtPDS) gene, whose mutation resulted in albino phenotypes. 16 Treatment of four independent transgenic lines with exogenous estradiol successfully induced 17 targeted mutagenesis in NtPDS. Sanger sequencing assay uncovered the presence of indel 18 mutations (nucleotides insertions or deletions) at the target site as expected, and at least two types 19 of mutations were identified for each line. Transgenic plants with mutated NtPDS gene after 20 estradiol treatment exhibited pale green or incomplete albino leaves. Moreover, the expression of 21 Cas9 in transgenic plants was strongly induced by estradiol treatment. Our results demonstrate 22 the efficacy of XVE-based CRISPR/Cas9 system in N. tabacum, and the system reported here 23 promises to be a useful approach for conditional genome editing, which would facilitate the study 24 of genes of interest, especially those developmentally important genes.25

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CRISPR–Cas9-mediated genome editing in apple and grapevine

Cited by 161 publications

References 43 publications

The state-of-the-art of grapevine biotechnology and new breeding technologies (NBTS)

The state-of-the-art of grapevine biotechnology and new breeding technologies (NBTS)

Development of Improved Fruit, Vegetable, and Ornamental Crops Using the CRISPR/Cas9 Genome Editing Technique

Targeted genome editing inNicotiana tabacumusing inducible CRISPR/Cas9 system

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