ArticlePlant biotechnology journal2025
Engineering a robust Cas12i3 variant-mediated wheat genome editing system.
Article in Plant biotechnology journal, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- Synergistic engineering of Casδ nuclease for robust genome editing.Journal of integrative plant biology · 2026Article
- Coupling of both a transactivation module and a double-stranded DNA-binding domain boosts Cas12i3 variant-based cytosine and adenine editing in plants.Journal of integrative plant biology · 2026Article
- Efficient CRISPR/Cas-SF01 genome editing tools with high editing efficiency in allotetraploid oilseed rape.Journal of integrative plant biology · 2026Article
- CasY7: An optimized Cas12i system for enhanced genome editing in monocot crops.Journal of integrative plant biology · 2026Article
- Developing a robust multiplex CRISPR/Cas12i3-5M system for trait stacking in soybean.Journal of integrative plant biology · 2026Article
- Optimized Cas-SF01 gene-editing toolbox shortens flowering timing in commercial maize inbred JING724.Journal of integrative plant biology · 2026Article
- Base editing in rice using nuclease-deactivated CRISPR/Cas-SF01.Journal of integrative plant biology · 2026Article
- Optimized tRNA processing and TREX2-SpCas9 fusion enable high-efficiency multiplex genome editing in plants.Plant communications · 2026Article
- Genome Editing Approaches in Flax (International journal of molecular sciences · 2026Review
- From Weed Evolution to Crop Design: A Computational Blueprint for a Novel, Synergistic Herbicide-Resistant Allele in Wheat.Plants (Basel, Switzerland) · 2026Article
- Wheat's Up with CRISPR-Cas-Current Advances, Obstacles and Perspectives.International journal of molecular sciences · 2026Review
- The Auxin Response FactorPlants (Basel, Switzerland) · 2026Article
- Beyond CRISPR/Cas9: emerging genome editing technologies for next-generation crop improvement.Molecular biology reports · 2026Review
- Engineering hypercompact IscB nucleases for efficient and versatile genome editing in rice.Genome biology · 2026Article
- Engineering the hypercompact miniature IscB-ωRNA systems for efficient rice genome editing.Plant communications · 2026Article
- exoCasMINI: A T5 exonuclease fused CRISPR-Cas12f system with enhanced gene editing efficiency.iScience · 2025Article
- Programmable genome engineering and gene modifications for plant biodesign.Plant communications · 2025Review
- Never-germinating Arabidopsis seeds with LbCas12a-induced mutations in 6 clade A type 2C protein phosphatase genes.Plant physiology · 2025Article
- A panomics-driven framework for the improvement of major food legume crops: advances, challenges, and future prospects.Horticulture research · 2025Article
- Genome Editing in Mouse Embryo Using the CRISPR/Cas12i3 System.International journal of molecular sciences · 2025Article
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7 authors.
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Abstract
Wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD) is one of the most important food crops in the world. CRISPR/Cas12i3, which belongs to the type V-I Cas system, has attracted extensive attention recently due to its smaller protein size and its less-restricted canonical 'TTN' protospacer adjacent motif (PAM). However, due to its relatively lower editing efficacy in plants and the hexaploidy complex nature of wheat, Cas12i3/Cas12i3-5M-mediated genome editing in wheat has not been documented yet. Here, we report the engineering of a robust Cas12i3-5M-mediated genome editing system in wheat through the fusion of T5 exonuclease (T5E) in combination with an optimised crRNA expression strategy (Opt). We first showed that fusion of T5E, rather than ExoI, to Cas12i3-5M increased the gene editing efficiencies by up to 1.34-fold and 3.87-fold, compared to Cas12i3-5M and Cas12i3 in HEK293T cells, respectively. However, its editing efficiency remains low in wheat. We then optimised the crRNA expression strategy and demonstrated that Opt-T5E-Cas12i3-5M could enhance the editing efficiency by 1.20- to 1.33-fold and 4.05- to 7.95-fold in wheat stable lines compared to Opt-Cas12i3-5M and Opt-Cas12i3, respectively, due to progressive 5'-end resection of the DNA strand at the cleavage site with increased deletion size. The Opt-T5E-Cas12i3-5M enabled an editing efficiency ranging from 60.71% to 90.00% across four endogenous target genes in stable lines of three elite Chinese wheat varieties. Together, the developed robust Opt-T5E-Cas12i3-5M system enriches wheat genome editing toolkits for either biological research or genetic improvement and may be extended to other important polyploidy crop species.
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