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CRISPR-Cas9-mediated genome editing in apple and grapevine
Yuriko Osakabe1, Zhenchang Liang2, Chong Ren2
1Faculty of Bioscience and Bioindustry, Tokushima University, Tokushima, Japan. osakabe.yuriko@tokushima-u.ac.jp.
Nature Protocols
|November 4, 2018
Summary
Scientists adapted CRISPR-Cas9 genome editing for apple and grapevine, enabling efficient DNA-free mutations. This tool accelerates crop improvement for desirable traits in fruit crops.
Area of Science:
- Agricultural Science
- Molecular Biology
- Genetics
Background:
- CRISPR-Cas9 technology and whole-genome sequencing have advanced targeted mutagenesis in crops.
- Efficient genome editing protocols are crucial for crop improvement in fruit species like apple and grapevine.
Purpose of the Study:
- To adapt and optimize CRISPR-Cas9 genome editing for apple and grapevine.
- To establish protocols for DNA-free targeted mutagenesis using both plasmid-mediated and ribonucleoprotein (RNP) delivery methods.
Main Methods:
- Developed and tested CRISPR-Cas9 protocols for apple and grapevine protoplasts.
- Utilized both plasmid-mediated genome editing and direct delivery of CRISPR-Cas9 ribonucleoproteins (RNPs).
- Verified targeted mutagenesis efficiency and regenerated edited plants.
Main Results:
- Achieved highly efficient targeted mutagenesis in apple and grapevine protoplasts.
- Demonstrated successful plant regeneration following plasmid-mediated CRISPR-Cas9 delivery.
- The RNP delivery method offers a faster alternative, with results in 2-3 weeks compared to >3 months for plasmid methods.
Conclusions:
- CRISPR-Cas9, via plasmid or RNP delivery, is effective for precise trait modulation in apple and grapevine.
- These adaptable protocols can accelerate genetic research and functional trait development in fruit crops.
- The DNA-free RNP method provides a rapid approach for genome editing in these species.
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