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Optimized prime editing in monocot plants using PlantPegDesigner and engineered plant prime editors (ePPEs) by Shuai Jin; Qiupeng Lin; Qiang Gao; Caixia Gao is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Optimized prime editing in monocot plants using PlantPegDesigner and engineered plant prime editors (ePPEs) about?

Prime editors (PEs), which can install desired base edits without donor DNA or double-strand breaks, have been used in plants and can, in principle, accelerate crop improvement and breeding. However, their editing efficiency in plants is generally low. Optimizing the prime editing guide RNA (pegRNA) by designing the sequence on the basis of melting temperature, using dual-pegRNAs and engineering PEs have all been shown to enhance PE efficiency. In addition, an automated pegRNA design platform, PlantPegDesigner, has been developed on the basis of rice prime editing experimental data. In this protocol, we present detailed protocols for designing and optimizing pegRNAs using PlantPegDesigner, constructing engineered plant PE vectors with enhanced editing efficiency for prime editing, evaluating prime editing efficiencies using a reporter system and comparing the effectiveness and byproducts of PEs by deep amplicon sequencing. Using this protocol, researchers can construct optimized pegRNAs for prime editing in 4-7 d and obtain prime-edited rice or wheat plants within 3 months.

Who reads Optimized prime editing in monocot plants using PlantPegDesigner and engineered plant prime editors (ePPEs)?

Author

Shuai Jin; Qiupeng Lin; Qiang Gao; Caixia Gao

Publisher

Springer Science and Business Media LLC

Published

2022

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)