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

Nat Biotechnol. 2021 Aug;39(8):923-927. doi: 10.1038/s41587-021-00868-w. Epub 2021 Mar 25.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems
  • Gene Editing / methods*
  • Oryza / genetics*
  • RNA, Guide / genetics*
  • RNA, Plant / genetics*


  • RNA, Guide
  • RNA, Plant