Book ChapterDOI
Genome Editing in Apple
Chikako Nishitani,Keishi Osakabe,Yuriko Osakabe +2 more
- pp 213-225
TLDR
This chapter will provide an overview of current studies of genome editing in apple and will discern and explore how this strategy will provide insights into molecular breeding technologies for genetic improvement of the apple.Abstract:
Genome editing with artificially engineered nucleases is an advanced molecular technology for pursuing precise and effective genetic engineering. In this technology, engineered nucleases induce DNA double-strand breaks at targeted sites in a genome, stimulating the DNA repair system in cells, thus enabling site-directed mutagenesis. Genome editing using CRISPR (clustered regularly interspaced short palindromic repeats)/CRISPR-associated protein9 (Cas9), originating from a defence system of prokaryotes, is a powerful technology that is now being widely utilized in molecular research studies, as well as in breeding programmes of various plant species, including fruit trees, to impart either novel or enhanced traits to established commercial cultivars or to new cultivars/genotypes. Recently, several reports have demonstrated successful apple genome editing and the introduction of important traits, such as those for early flowering and reduced fire blight susceptibility, to popular commercial cultivars, such as ‘Gala’ and ‘Golden Delicious’. It is important to point out that these reports reveal that such genome-edited/mutant apple plants or cell lines do not carry foreign genes. Nevertheless, during the process of precise genome editing, the coexistence of various types of mutations referred to as “mosaic mutations” and off-target effects are major concerns. Therefore, to minimize such effects, selection of target sequences and estimation of off-target effects for CRISPR/Cas9 has been developed for many organisms, and these have also been employed for apple by using in silico analysis based on genome information. On the other hand, apple genome heterozygosity has led to difficulties in genome editing, as the complex genome of apple precludes the use of some of these basic techniques for genome editing. Therefore, further studies focused on genome information and culture techniques tailored for apple are needed. It will be highly critical for each apple cultivar in developing precise and efficient genome editing for apple. This chapter will provide an overview of current studies of genome editing in apple and will discern and explore how this strategy will provide insights into molecular breeding technologies for genetic improvement of the apple.read more
Citations
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Journal ArticleDOI
Breeding and genetics of disease resistance in temperate fruit trees: challenges and new opportunities
Awais Khan,Schuyler S. Korban +1 more
Journal ArticleDOI
Spontaneous, Artificial, and Genome Editing-Mediated Mutations in Prunus
Ángela S. Prudencio,Sama Rahimi Devin,S. Mahdavi,Pedro J. Martínez-García,Juan A. Salazar,Pedro Martínez-Gómez +5 more
TL;DR: This paper reviewed the most relevant mutations for breeding traits in Prunus species such as flowering time, self-compatibility, fruit quality, and disease tolerance, including new molecular perspectives in the present postgenomic era including CRISPR/Cas9 and TALEN editing technologies.
References
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Journal ArticleDOI
Mosaicism in CRISPR/Cas9-mediated genome editing
TL;DR: By developing ways to overcome mosaic mutations when usingCRISPR/Cas9, genotyping for germline gene disruptions should become more reliable and pave the way for using the CRISPR technology in the research and clinical applications where mosaicism is an issue.
Journal ArticleDOI
Sequence features associated with the cleavage efficiency of CRISPR/Cas9 system.
TL;DR: Important parameters for the design of sgRNAs with high on-target efficiencies are revealed, especially in the context of high throughput applications, including CRISPR-Cas9 applications.
Journal ArticleDOI
Efficient Targeted Mutagenesis in Apple and First Time Edition of Pear Using the CRISPR-Cas9 System.
Aurélie Charrier,Emilie Vergne,Nicolas Dousset,Andréa Richer,Aurélien Petiteau,Elisabeth Chevreau +5 more
TL;DR: The overall results indicate that, despite the frequent occurrence of chimerism, the CRISPR-Cas 9 system is a powerful and precise method to induce targeted mutagenesis in the first generation of apple and pear transgenic lines.
Journal ArticleDOI
CRISPR-Cas9-mediated genome editing in apple and grapevine.
Yuriko Osakabe,Zhenchang Liang,Chong Ren,Chikako Nishitani,Keishi Osakabe,Masato Wada,Sadao Komori,Mickael Malnoy,Riccardo Velasco,Michele Poli,Minhee Jung,Ok Jae Koo,Roberto Viola,Chidananda Nagamangala Kanchiswamy +13 more
TL;DR: This protocol provides a stepwise protocol for the design and transfer of CRISPR–Cas9 components to apple and grapevine protoplasts, followed by verification of highly efficient targeted mutagenesis, and regeneration of plants following the plasmid-mediated delivery of components.
Journal ArticleDOI
Optimized paired-sgRNA/Cas9 cloning and expression cassette triggers high-efficiency multiplex genome editing in kiwifruit.
TL;DR: It is concluded that the PTG/Cas9 system is a more powerful system than the traditional CRISPR/cas9 system for kiwifruit genome editing, which provides valuable clues for optimizing CRISpr/Cas 9 editing system in other plants.