ArticlePlant & cell physiology2016
Precision Targeted Mutagenesis via Cas9 Paired Nickases in Rice.
Article in Plant & cell physiology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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Who cites it
40 citing papers in PubMed, 97 citations in OpenAlex.
- Targeted mutagenesis and base editing using engineeredPlant biotechnology (Tokyo, Japan) · 2026Article
- Versatile genome editing using Type I-E CRISPR-Cas3 in rice.Plant & cell physiology · 2026Article
- A 65-kb deletion survey identifies a distalProceedings of the National Academy of Sciences of the United States of America · 2025Article
- Recent developments, challenges and opportunities in genome editing for crop science from a societal perspective.Frontiers in genome editing · 2025Review
- CRISPR/Cas9-mediated mutagenesis of phytoene desaturase in pigeonpea and groundnut.Functional & integrative genomics · 2024Article
- Prime editing: Mechanism insight and recent applications in plants.Plant biotechnology journal · 2024Review
- CRISPR/Cas as a Genome-Editing Technique in Fruit Tree Breeding.International journal of molecular sciences · 2023Review
- Targeted mutagenesis with sequence-specific nucleases for accelerated improvement of polyploid crops: Progress, challenges, and prospects.The plant genome · 2023Review
- CRISPR/Cas9: an advanced platform for root and tuber crops improvement.Frontiers in genome editing · 2023Review
- Recent Advances in Improving Gene-Editing Specificity through CRISPR-Cas9 Nuclease Engineering.Cells · 2022Review
- CRISPR-Based Genome Editing: Advancements and Opportunities for Rice Improvement.International journal of molecular sciences · 2022Review
- Different DNA repair pathways are involved in single-strand break-induced genomic changes in plants.The Plant cell · 2021Article
- Optimized design parameters for CRISPR Cas9 and Cas12a homology-directed repair.Scientific reports · 2021Article
- Induced Genetic Variations in Fruit Trees Using New Breeding Tools: Food Security and Climate Resilience.Plants (Basel, Switzerland) · 2021Review
- Advanced domestication: harnessing the precision of gene editing in crop breeding.Plant biotechnology journal · 2021Review
- Approach for in vivo delivery of CRISPR/Cas system: a recent update and future prospect.Cellular and molecular life sciences : CMLS · 2021Review
- Applications of CRISPR/Cas to Improve Crop Disease Resistance: Beyond Inactivation of Susceptibility Factors.iScience · 2020Review
- Improving the genome editing efficiency of CRISPR/Cas9 in Arabidopsis and Medicago truncatula.Planta · 2020Article
- Review
- How to start your monocot CRISPR/Cas project: plasmid design, efficiency detection, and offspring analysis.Rice (New York, N.Y.) · 2020Review
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recent reports of CRISPR- (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) mediated heritable mutagenesis in plants highlight the need for accuracy of the mutagenesis directed by this system. Off-target mutations are an important issue when considering functional gene analysis, as well as the molecular breeding of crop plants with large genome size, i.e. with many duplicated genes, and where the whole-genome sequence is still lacking. In mammals, off-target mutations can be suppressed by using Cas9 paired nickases together with paired guide RNAs (gRNAs). However, the performance of Cas9 paired nickases has not yet been fully assessed in plants. Here, we analyzed on- and off-target mutation frequency in rice calli and regenerated plants using Cas9 nuclease or Cas9 nickase with paired gRNAs. When Cas9 paired nickases were used, off-target mutations were fully suppressed in rice calli and regenerated plants. However, on-target mutation frequency also decreased compared with that induced by the Cas9 paired nucleases system. Since the gRNA sequence determines specific binding of Cas9 protein-gRNA ribonucleoproteins at the targeted sequence, the on-target mutation frequency of Cas9 paired nickases depends on the design of paired gRNAs. Our results suggest that a combination of gRNAs that can induce mutations at high efficiency with Cas9 nuclease should be used together with Cas9 nickase. Furthermore, we confirmed that a combination of gRNAs containing a one nucleotide (1 nt) mismatch toward the target sequence could not induce mutations when expressed with Cas9 nickase. Our results clearly show the effectiveness of Cas9 paired nickases in delivering on-target specific mutations.
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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.