Spelling Changes and Fluorescent Tagging With Prime Editing Vectors for Plants

Area of Science:

• Molecular Biology
• Plant Science
• Biotechnology

Background:

• CRISPR-Cas systems offer precise genome editing capabilities.
• Prime editing, a derivative of CRISPR-Cas, allows targeted DNA modifications via RNA templates.
• Previous plant prime editing studies reported limited insertion sizes.

Purpose of the Study:

• To develop and validate user-friendly prime editing vectors for both dicot and monocot plants.
• To demonstrate enhanced prime editing capabilities, specifically larger DNA insertions.
• To enable advanced applications like fluorescent tagging in plants.

Main Methods:

• Construction of novel prime editing vectors compatible with diverse plant species.
• Validation of prime editing efficiency in model plants: *Nicotiana benthamiana*, rice, and *Arabidopsis thaliana*.
• Utilizing prime editing to achieve a significant DNA insertion for split-GFP tagging.

Main Results:

• Development of versatile prime editing vectors applicable to monocots and dicots.
• Successful demonstration of prime editing in *N. benthamiana*, rice, and *Arabidopsis*.
• Achieved a 66 bp DNA insertion, substantially larger than previously reported in plants, enabling split-GFP fluorescent protein complementation.

Conclusions:

• The developed prime editing vectors are effective tools for precise genome engineering in a wide range of plant species.
• This advancement significantly expands the utility of prime editing in plants, particularly for generating larger insertions.
• The successful split-GFP tagging showcases the potential for advanced molecular and genetic applications in plants using prime editing.