3.8 Article

Spelling Changes and Fluorescent Tagging With Prime Editing Vectors for Plants

Journal

FRONTIERS IN GENOME EDITING
Volume 3, Issue -, Pages -

Publisher

FRONTIERS MEDIA SA
DOI: 10.3389/fgeed.2021.617553

Keywords

prime editing; plant genome editing; fluorescent tagging; split GFP; Oryza sativa; Arabidopsis; Nicotiana benthamiana

Funding

  1. National Science Foundation [IOS-1444511, IOS-1546625]
  2. National Institute of Food and Agriculture of the U.S. Department of Agriculture [2018-67011-28025]
  3. Indo-U.S. Science and Technology Forum, Government of India
  4. Manisa Celal Bayar University Scientific Research Projects funds [2017-113]
  5. National Institutes of Health [S10RR025502]

Ask authors/readers for more resources

Prime editing is an improved version of the CRISPR-Cas system, utilizing a fusion protein and guide RNA to achieve specified substitutions, insertions, and deletions in the target DNA. Recent studies have demonstrated successful Prime editing in rice and wheat, including significant insertions.
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.

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