A flanking-nicks prime editor (FLICK-PE) system to boost prime editing in dicots

Area of Science:

• Plant Biotechnology
• Genome Editing
• Molecular Biology

Background:

• Prime editing (PE) offers precise genome modification but faces challenges in dicot plants.
• Existing PE systems have limitations in efficiency for certain crop species.

Purpose of the Study:

• To develop an optimized prime editing system for enhanced efficiency in dicot plants.
• To engineer desirable traits, such as herbicide resistance, in agriculturally important crops.

Main Methods:

• Development of the Flanking-Nicks Prime Editor (FLICK-PE) system.
• Optimization of prime editing by incorporating flanking nicking sgRNAs.
• Application of FLICK-PE for introducing specific mutations in soybean (EPSPS1a) and tobacco.

Main Results:

• FLICK-PE demonstrated a significant increase in intended-editing efficiency in soybean (15.7-fold vs. PE2, 2.2-fold vs. PE3).
• Engineered glyphosate resistance in soybean with high editing efficiency (21.1%) and stable trait inheritance.
• FLICK-PE proved effective in tobacco, indicating broad applicability.

Conclusions:

• FLICK-PE represents a substantial advancement for precision genome editing in dicot crops.
• This technology facilitates rapid trait development and precision breeding in agriculture.
• FLICK-PE holds promise for accelerating the creation of improved crop varieties with enhanced traits.