ArticleJournal of experimental botany2023
Improved prime editing allows for routine predictable gene editing in Physcomitrium patens.
Article in Journal of experimental botany, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 19 citations in OpenAlex.
- Transgene-free genome editing in plants.aBIOTECH · 2026Review
- Prime editing: evolution of CRISPR-Cas system for a robust next-generation genome editing in plants.Planta · 2026Review
- The Importance of thePlants (Basel, Switzerland) · 2026Review
- tRNA-based polycistronic CRISPR/Cas9 system boosts efficiency of multi-gene deletion in the moss Physcomitrella.Plant methods · 2026Article
- Research progress of base editing and prime editing tools based on the CRISPR/Cas system.Molecular therapy. Nucleic acids · 2025Review
- Moss BRCA2 lacking the canonical DNA-binding domain promotes homologous recombination and binds to DNA.Nucleic acids research · 2025Article
- Emerging trends in transgene-free crop development: insights into genome editing and its regulatory overview.Plant molecular biology · 2025Review
- Review
- Emerging applications of gene editing technologies for the development of climate-resilient crops.Frontiers in genome editing · 2025Review
- A simple and efficient system for evaluating plant genome editing efficiency and its application in optimizing the ISAam1 TnpB nuclease.Frontiers in plant science · 2025Article
- Isolation and Transfection of Nicotiana benthamiana Mesophyll Protoplasts.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Editing VvDXS1 for the creation of muscat flavour in Vitis vinifera cv. Scarlet Royal.Plant biotechnology journal · 2024Article
- Article
- The SpRY Cas9 variant release the PAM sequence constraint for genome editing in the model plant Physcomitrium patens.Transgenic research · 2024Article
- Prime editing: Mechanism insight and recent applications in plants.Plant biotechnology journal · 2024Review
- Advances in plant molecular biology: towards new challenges.Journal of experimental botany · 2023Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Efficient and precise gene editing is the gold standard of any reverse genetic study. The recently developed prime editing approach, a modified CRISPR/Cas9 [clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein] editing method, has reached the precision goal but its editing rate can be improved. We present an improved methodology that allows for routine prime editing in the model plant Physcomitrium patens, whilst exploring potential new prime editing improvements. Using a standardized protoplast transfection procedure, multiple prime editing guide RNA (pegRNA) structural and prime editor variants were evaluated targeting the APT reporter gene through direct plant selection. Together, enhancements of expression of the prime editor, modifications of the 3' extension of the pegRNA, and the addition of synonymous mutation in the reverse transcriptase template sequence of the pegRNA dramatically improve the editing rate without affecting the quality of the edits. Furthermore, we show that prime editing is amenable to edit a gene of interest through indirect selection, as demonstrated by the generation of a Ppdek10 mutant. Additionally, we determine that a plant retrotransposon reverse transcriptase enables prime editing. Finally, we show for the first time the possibility of performing prime editing with two independently coded peptides.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.