Spelling Changes and Fluorescent Tagging With Prime Editing Vectors for Plants

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Wang L., Kaya H. B., Zhang N., Rai R., Willmann M. R., Carpenter S. C. D., ...More

Frontiers in Genome Editing, vol.3, 2021 (Scopus) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 3
  • Publication Date: 2021
  • Doi Number: 10.3389/fgeed.2021.617553
  • Journal Name: Frontiers in Genome Editing
  • Journal Indexes: Scopus, Directory of Open Access Journals
  • Keywords: prime editing, plant genome editing, fluorescent tagging, split GFP, Oryza sativa, Arabidopsis, Nicotiana benthamiana
  • Open Archive Collection: AVESIS Open Access Collection
  • Manisa Celal Bayar University Affiliated: No

Abstract

Prime editing is an adaptation of the CRISPR-Cas system that uses a Cas9(H840A)-reverse transcriptase fusion and a guide RNA amended with template and primer binding site sequences to achieve RNA-templated conversion of the target DNA, allowing specified substitutions, insertions, and deletions. In the first report of prime editing in plants, a variety of edits in rice and wheat were described, including insertions up to 15 bp. Several studies in rice quickly followed, but none reported a larger insertion. Here, we report easy-to-use vectors for prime editing in dicots as well as monocots, their validation in Nicotiana benthamiana, rice, and Arabidopsis, and an insertion of 66 bp that enabled split-GFP fluorescent tagging.