High-efficiency prime editing with optimized, paired pegRNAs in plants

Prime editing (PE) applications are limited by low editing efficiency. Here we show that designing prime binding sites with a melting temperature of 30 °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...

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Veröffentlicht in:	Nature biotechnology 2021-08, Vol.39 (8), p.923-927
Hauptverfasser:	Lin, Qiupeng, Jin, Shuai, Zong, Yuan, Yu, Hong, Zhu, Zixu, Liu, Guanwen, Kou, Liquan, Wang, Yanpeng, Qiu, Jin-Long, Li, Jiayang, Gao, Caixia
Format:	Artikel
Sprache:	eng
Schlagworte:	631/449/2491 631/61/447 Agriculture Analysis Applications programs Binding sites Bioinformatics Biomedical and Life Sciences Biomedical Engineering/Biotechnology Biomedicine Biotechnology Brief Communication CRISPR-Cas Systems Editing Efficiency Gene Editing - methods Genetic aspects Genomics Life Sciences Melt temperature Oryza - genetics Physiological aspects Plant genetics Rice RNA, Guide, CRISPR-Cas Systems RNA, Plant - genetics
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Beschreibung
Zusammenfassung:	Prime editing (PE) applications are limited by low editing efficiency. Here we show that designing prime binding sites with a melting temperature of 30 °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.
ISSN:	1087-0156 1546-1696
DOI:	10.1038/s41587-021-00868-w