ArticleJournal of advanced research2025
Developing a multi-modular assembled prime editing (mPE) system improved precise multi-base insertion efficiency in dicots.
Article in Journal of advanced research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers, 1 of them a synthesis that pooled it.
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Who cites it
6 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Pooled it
- Prime editing: evolution of CRISPR-Cas system for a robust next-generation genome editing in plants.Planta · 2026Review
- Efficient prime editors for heritable multiplex precision genome editing in soybean.Nature plants · 2026Article
- Uncleavable fusion of Csy4 with prime editors for low Csy4 toxicity and highly efficient prime editing in Arabidopsis.Plant physiology · 2026Article
- Development of an ultra-efficient prime editing system in tomato.Nature communications · 2026Article
- Establishment of a TRV-based virus-induced gene silencing system using the vacuum infiltration in Tartary buckwheat.Plant cell reports · 2025Article
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3 authors.
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Abstract
introductionThe Prime Editing (PE) system is a precise and versatile genome editing tool with great potential in plant breeding and plant synthetic biology. However, low PE efficiency severely restricts its application, especially in dicots. PE can introduce small tags to trace target protein or cis-element to regulate gene transcription which is an expertise superior to other gene editing tools. Owing to low efficiency, PE adaption in stably transformed Arabidopsis is lacking.
objectivesThis study aimed to investigate the issue of low PE efficiency in dicots and develop systematic solutions to improve it. Currently, PE in dicots is undetectable and inconsistent, and this study seeks to address it. Split PE into several parts showed better performance in some target sites in mammal cells. We plan to discover the optimal split PE combination in dicot.
methodsWe conducted large-scale transformation experiments in dicot model plants Arabidopsis thaliana (At) and Nicotiana benthamiana (Nb) by Agrobacterium-mediated transformation with deep amplicon sequencing (0.2-0.5 million clean total reads).
resultsThe editing efficiency decreased upon using a fused reverse transcriptase (RT) or an extended pegRNA separately and further decreased dramatically when these were used together. With the help of the pol II strategy to express PE gRNA (pegRNA), we named the most effective split PE combination as a multi-modular assembled prime editing system (mPE). mPE exhibited improved precise editing efficiency on most gene sites with various editing types, ranging from 1.3-fold to 1288.5-fold and achieved PE on some sites that could not be edited by original PE2. Especially, mPE showed superiority for multi-base insertion with an average improvement of 197.9-fold.
conclusionThe original PE architecture strongly inhibited the cleavage activity of Cas9. Split PE improved PE efficiency extensively and was in favor of introducing small insertions in dicot plants, indicating that different PE variants might have their own expertise.
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