ArticleThe Plant journal : for cell and molecular biology2017
High-efficiency gene targeting in hexaploid wheat using DNA replicons and CRISPR/Cas9.
Article in The Plant journal : for cell and molecular biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 167 papers.
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Who cites it
167 citing papers in PubMed, 354 citations in OpenAlex.
- Unlocking the future of crop improvement with crop wild relatives.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2026Review
- Transposase-Assisted Donor Tethering Boosts Large-Fragment HDR in Plants.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Genome Editing Approaches in Flax (International journal of molecular sciences · 2026Review
- Wheat's Up with CRISPR-Cas-Current Advances, Obstacles and Perspectives.International journal of molecular sciences · 2026Review
- Plant viruses as next-generation vectors for transgene-free genome editing, gene regulation, and rapid crop improvement.Plant molecular biology · 2026Review
- From Gene Knockouts to Genome Remodeling: Large DNA Fragment Deletion Technologies in Plants.Plants (Basel, Switzerland) · 2026Review
- Development of Modular Geminivirus-Based Vectors for High Cargo Expression and Gene Targeting in Plants.Plant biotechnology journal · 2026Article
- Geminiviral-CR-gRNA expressed in cowpea efficiently edited MYMV and MYMIV genome to provide resistance against cowpea yellow mosaic disease without hampering plant growth and yield.BMC plant biology · 2026Article
- Synthetic Biology of Plants and Microbes for Agriculture, Environment, and Future Applications.Chemical reviews · 2026Review
- Geminivirus-derived replicons: assessment for transient GFP expression in tobacco and tomato.Frontiers in plant science · 2026Article
- CRISPR-Cas technologies for precision genome editing in plants: advances, applications, and future perspectives.Frontiers in plant science · 2026Review
- Rye sprout as a host for the rapid transient production of a recombinant protein using a geminivirus DNA-containing expression vector.Plant biotechnology (Tokyo, Japan) · 2025Article
- Article
- CRISPR/Cas-mediated genome editing: playing a versatile role in mitigating the challenges of sustainable rice improvement.3 Biotech · 2025Review
- Bioluminescence-Driven Optimization of Geminivirus-Based Vectors as Tools for Plant Biotechnology.ACS synthetic biology · 2025Article
- Miniature enOsCas12f1 Enables Targeted Genome Editing in Rice.Plants (Basel, Switzerland) · 2025Article
- CRISPR/Cas genome editing in soybean: challenges and new insights to overcome existing bottlenecks.Journal of advanced research · 2025Review
- Cas9-Rep fusion tethers donor DNA in vivo and boosts the efficiency of HDR-mediated genome editing.Plant biotechnology journal · 2025Article
- CRISPR-Cas applications in agriculture and plant research.Nature reviews. Molecular cell biology · 2025Review
- Simplex and multiplex CRISPR/Cas9-mediated knockout of grain protease inhibitors in model and commercial barley improves hydrolysis of barley and soy storage proteins.Plant biotechnology journal · 2025Article
107 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
11 authors at 4 institutions in 3 countries.
Funding
Abstract
The ability to edit plant genomes through gene targeting (GT) requires efficient methods to deliver both sequence-specific nucleases (SSNs) and repair templates to plant cells. This is typically achieved using Agrobacterium T-DNA, biolistics or by stably integrating nuclease-encoding cassettes and repair templates into the plant genome. In dicotyledonous plants, such as Nicotinana tabacum (tobacco) and Solanum lycopersicum (tomato), greater than 10-fold enhancements in GT frequencies have been achieved using DNA virus-based replicons. These replicons transiently amplify to high copy numbers in plant cells to deliver abundant SSNs and repair templates to achieve targeted gene modification. In the present work, we developed a replicon-based system for genome engineering of cereal crops using a deconstructed version of the wheat dwarf virus (WDV). In wheat cells, the replicons achieve a 110-fold increase in expression of a reporter gene relative to non-replicating controls. Furthermore, replicons carrying CRISPR/Cas9 nucleases and repair templates achieved GT at an endogenous ubiquitin locus at frequencies 12-fold greater than non-viral delivery methods. The use of a strong promoter to express Cas9 was critical to attain these high GT frequencies. We also demonstrate gene-targeted integration by homologous recombination (HR) in all three of the homoeoalleles (A, B and D) of the hexaploid wheat genome, and we show that with the WDV replicons, multiplexed GT within the same wheat cell can be achieved at frequencies of ~1%. In conclusion, high frequencies of GT using WDV-based DNA replicons will make it possible to edit complex cereal genomes without the need to integrate GT reagents into the genome.
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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.