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Optimization of Prime Editing in Rice, Peanut, Chickpea, and Cowpea Protoplasts by Restoration of GFP Activity
| Content Provider | MDPI |
|---|---|
| Author | Biswas, Sudip Bridgeland, Aya Irum, Samra Thomson, Michael J. Septiningsih, Endang M. |
| Copyright Year | 2022 |
| Description | Precise editing of the plant genome has long been desired for functional genomic research and crop breeding. Prime editing is a newly developed precise editing technology based on CRISPR-Cas9, which uses an engineered reverse transcriptase (RT), a catalytically impaired Cas9 endonuclease (nCas9), and a prime editing guide RNA (pegRNA). In addition, prime editing has a wider range of editing types than base editing and can produce nearly all types of edits. Although prime editing was first established in human cells, it has recently been applied to plants. As a relatively new technique, optimization will be needed to increase the editing efficiency in different crops. In this study, we successfully edited a mutant GFP in rice, peanut, chickpea, and cowpea protoplasts. In rice, up to 16 times higher editing efficiency was achieved with a dual pegRNA than the single pegRNA containing vectors. Edited-mutant GFP protoplasts have also been obtained in peanut, chickpea, and cowpea after transformation with the dual pegRNA vectors, albeit with much lower editing efficiency than in rice, ranging from 0.2% to 0.5%. These initial results promise to expedite the application of prime editing in legume breeding programs to accelerate crop improvement. |
| Starting Page | 9809 |
| e-ISSN | 14220067 |
| DOI | 10.3390/ijms23179809 |
| Journal | International Journal of Molecular Sciences |
| Issue Number | 17 |
| Volume Number | 23 |
| Language | English |
| Publisher | MDPI |
| Publisher Date | 2022-08-29 |
| Access Restriction | Open |
| Subject Keyword | International Journal of Molecular Sciences Plant Sciences Prime Editing Crispr-cas9 Dual Pegrna Mutant Gfp Legume Editing Efficiency |
| Content Type | Text |
| Resource Type | Article |
