Journal
NATURE BIOTECHNOLOGY
Volume 39, Issue 8, Pages 923-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41587-021-00868-w
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Funding
- National Transgenic Science and Technology Program of China [2019ZX08010-001, 2019ZX08010-003]
- Strategic Priority Research Program of the Chinese Academy of Sciences (Precision Seed Design and Breeding) [XDA24020310, XDA24030504]
- National Natural Science Foundation of China [31788103, 32001060]
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Designing prime binding sites with a certain melting temperature and using two pegRNAs can substantially enhance the efficiency of prime editing.
Prime editing (PE) applications are limited by low editing efficiency. Here we show that designing prime binding sites with a melting temperature of 30 degrees C leads to optimal performance in rice and that using two prime editing guide (peg) RNAs in trans encoding the same edits substantially enhances PE efficiency. Together, these approaches boost PE efficiency from 2.9-fold to 17.4-fold. Optimal pegRNAs or pegRNA pairs can be designed with our web application, PlantPegDesigner. Improved guide RNAs enhance the efficiency of prime editing.
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