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CRISPR/Cas precision: do we need to worry about off-targeting in plants?
Florian Hahn1, Vladimir Nekrasov2
1Plant Sciences Department, Rothamsted Research, Harpenden, AL5 2JQ, UK.
Plant Cell Reports
|November 15, 2018
Summary
CRISPR/Cas gene editing in plants can cause unintended mutations. This review details strategies like using precise nucleases (e.g., Cas12a) and ribonucleoproteins (RNPs) to improve accuracy and avoid off-target effects.
Area of Science:
- Plant Science
- Molecular Biology
- Biotechnology
Background:
- CRISPR/Cas technology is a powerful tool for targeted genome modification in plants.
- Off-target mutations are a significant concern, frequently reported in mammalian systems and also relevant in plants.
Purpose of the Study:
- To summarize current knowledge on CRISPR/Cas precision in plants.
- To outline strategies for minimizing off-target mutations and unintended on-target changes.
Main Methods:
- Review of existing literature on CRISPR/Cas precision and off-target effects in plants.
- Discussion of advanced CRISPR/Cas nucleases (e.g., Cas12a, SpCas9-HF) and delivery methods (ribonucleoproteins).
Main Results:
- Higher precision nucleases and ribonucleoprotein delivery can reduce off-target mutations.
- Careful target sequence selection is the most critical factor for minimizing CRISPR/Cas errors.
Conclusions:
- Implementing precise nucleases, RNPs, and rigorous target site selection are key to enhancing CRISPR/Cas accuracy in plants.
- Available online tools and experimental guidance can aid in selecting optimal target sequences.
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