Boosting genome editing in plants with single transcript unit surrogate reporter systems.

Area of Science:

• Plant genomics
• Genome editing technologies
• Crop improvement

Background:

• CRISPR-Cas genome editing offers significant potential for plant science and crop development.
• Identifying successful genome editing events can be challenging due to low editing efficiency.

Purpose of the Study:

• To develop and validate novel reporter systems for enhanced selection of genome-edited plants.
• To establish a direct correlation between reporter gene editing and endogenous gene editing.

Main Methods:

• Introduction of multiple single transcript unit surrogate reporter (STU-SR) systems.
• Utilizing strategies like single-strand annealing (SSA) and base editing for reporter gene restoration.
• Employing STU-SR systems with Cas9 nuclease, cytosine base editors, and adenine base editors.

Main Results:

• STU-SR systems effectively enhanced genome editing event identification in rice (monocot).
• Demonstrated successful application in both Cas9 nuclease-based mutagenesis and base editing (cytosine and adenine).
• Showcased compatibility with Cas9 variants (e.g., SpRY) and improved editing in Brassica oleracea (dicot).

Conclusions:

• Developed highly efficient and versatile STU-SR systems for enriching genome-edited plants.
• These systems provide a robust method for selecting edited individuals across different plant species and editing types.