Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize

Genome Biol. 2020 Oct 6;21(1):257. doi: 10.1186/s13059-020-02170-5.

Abstract

Prime editing is a novel and universal CRISPR/Cas-derived precision genome-editing technology that has been recently developed. However, low efficiency of prime editing has been shown in transgenic rice lines. We hypothesize that enhancing pegRNA expression could improve prime-editing efficiency. In this report, we describe two strategies for enhancing pegRNA expression. We construct a prime editing vector harboring two pegRNA variants for W542L and S621I double mutations in ZmALS1 and ZmALS2. Compared with previous reports in rice, we achieve much higher prime-editing efficiency in maize. Our results are inspiring and provide a direction for the optimization of plant prime editors.

Keywords: ALS; CRISPR/Cas9; Herbicide resistance; Maize; PE2; Prime editing; pegRNA.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetolactate Synthase / genetics*
  • Gene Editing / methods*
  • Gene Editing / statistics & numerical data
  • Genetic Vectors
  • Mutagenesis, Site-Directed / methods*
  • Plants, Genetically Modified
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • RNA, Guide, CRISPR-Cas Systems / metabolism*
  • Zea mays / enzymology
  • Zea mays / genetics*

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • Acetolactate Synthase