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Virus-Mediated Genome Editing in Plants Using the CRISPR/Cas9 System
Ahmed Mahas1, Zahir Ali1, Manal Tashkandi1
1Laboratory for Genome Engineering, Division of Environmental and Biological Sciences and Engineering, 4700 King Abdullah University of Science and Technology, Thuwal, Saudi Arabia.
Methods in Molecular Biology (Clifton, N.J.)
|January 6, 2019
Summary
Researchers developed a Tobacco rattle virus (TRV)-mediated system for efficient CRISPR/Cas9 genome editing in plants. This method delivers editing reagents systemically, enabling targeted plant genome modification without tissue culture.
Area of Science:
- Plant science
- Molecular biology
- Genetics
Background:
- Targeted plant genome modification is crucial for agricultural innovation.
- CRISPR/Cas9 is a popular genome-editing tool, but efficient delivery of reagents in plants remains challenging.
- In planta genome engineering requires overcoming delivery hurdles to modify desired plant tissues.
Purpose of the Study:
- To develop an efficient method for delivering genome engineering reagents in plants.
- To establish a Tobacco rattle virus (TRV)-mediated system for in planta genome modification.
- To bypass the need for plant transformation and tissue culture in genome engineering.
Main Methods:
- Development of a TRV-mediated genome engineering system for Nicotiana benthamiana.
- Delivery of Cas9 protein and guide RNAs into plant cells via TRV.
- Recovery of progeny plants with desired genomic modifications.
Main Results:
- The TRV-based system enables efficient, systemic delivery of genome engineering reagents in N. benthamiana.
- This approach facilitates targeted genome modification in all plant parts.
- Progeny plants with desired modifications were recovered without transformation or tissue culture.
Conclusions:
- TRV-mediated genome engineering offers a simple and efficient platform for in planta modification.
- This system expands the utility of CRISPR/Cas9 for targeted genome editing in plants.
- The protocol can be adapted for other plant species and potentially other RNA virus vectors.
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