An efficient CRISPR-Cas12a promoter editing system for crop improvement

Area of Science:

• Agricultural Science
• Molecular Biology
• Genetics

Background:

• Promoter editing is a novel strategy for generating quantitative trait variation (QTV) in crops.
• An efficient and versatile promoter editing system is crucial for crop improvement.

Purpose of the Study:

• To develop and validate an efficient CRISPR-Cas12a-based promoter editing system for generating QTV in crops.
• To demonstrate the system's utility in rice for traits like grain quality and plant architecture.

Main Methods:

• Developed the CRISPR-Cas12a promoter editing (CAPE) system, integrating a promoter key-region estimating model.
• Applied CAPE for multiplexed and singular editing in rice, targeting genes like OsGBSS1, OsGS3, and OsD18.
• Conducted field trials to evaluate the performance of edited lines.

Main Results:

• CAPE successfully generated QTV continuums for grain starch content and size in rice.
• Promoter editing of OsD18 resulted in semidwarf rice lines with a QTV continuum for plant height.
• OsD18 edited lines exhibited comparable yield and lodging resistance to Green Revolution mutants.

Conclusions:

• The CAPE system provides an efficient pipeline for producing useful QTV continuums in crops.
• Promoter editing of OsD18 generates a quantitative Green Revolution trait, offering a new avenue for crop breeding.
• This work establishes a robust platform for targeted crop trait improvement through promoter editing.