ArticlePlant molecular biology2018
Potential high-frequency off-target mutagenesis induced by CRISPR/Cas9 in Arabidopsis and its prevention.
Article in Plant molecular biology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 83 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
83 citing papers in PubMed, 194 citations in OpenAlex.
- Dexamethasone-inducible LhGR/pOp system: simple and flexible spatiotemporal control of gene expression.Planta · 2026Review
- Nanotechnology-Enabled CRISPR Delivery: Emerging Opportunities in Agriculture and Forest Biotechnology.Plants (Basel, Switzerland) · 2026Review
- Protoplast-Based Functional Genomics and Genome Editing: Progress, Challenges and Applications.Plant, cell & environment · 2026Review
- Multiplex gene editing drives revolution in crop breeding: overlaid editing of multiple genes and customization of complex traits.Advanced biotechnology · 2026Review
- Versatile Applications of CRISPR-Based Programmable T-DNA Integration in Plants.Plant biotechnology journal · 2025Article
- Targeted mutagenesis ofaBIOTECH · 2025Article
- Mechanistic insights into Cas13d enzymes from cryo-EM structures of CasRx and DjCas13d.Nucleic acids research · 2025Article
- Ribosome biogenesis in plants requires the nuclear envelope and mitochondria localized OPENER complex.Nature communications · 2025Article
- RNA-Seq Identification of Peanut Callus-Specific Promoters and Evaluation of Base-Editing Efficiency.Plants (Basel, Switzerland) · 2025Article
- Suitability of Real-Time PCR Methods for New Genomic Technique Detection in the Context of the European Regulations: A Case Study in Arabidopsis.International journal of molecular sciences · 2025Article
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- Natural variation in TaERF-A1 confers semi-dwarf and lodging-resistant plant architecture in wheat.Plant communications · 2025Article
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- Editing theInternational journal of molecular sciences · 2025Article
- Recent developments, challenges and opportunities in genome editing for crop science from a societal perspective.Frontiers in genome editing · 2025Review
- From pandemics to preparedness: harnessing AI, CRISPR, and synthetic biology to counter biosecurity threats.Frontiers in public health · 2025Review
- Cas12a-mediated gene targeting by sequential transformation strategy in Arabidopsis thaliana.BMC plant biology · 2024Article
- Opportunity for genome engineering to enhance phosphate homeostasis in crops.Physiology and molecular biology of plants : an international journal of functional plant biology · 2024Review
- A dual-function selection system enables positive selection of multigene CRISPR mutants and negative selection ofaBIOTECH · 2024Article
- Genetic dissection of mutagenic repair and T-DNA capture at CRISPR-induced DNA breaks inPNAS nexus · 2024Article
23 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
key messageWe present novel observations of high-specificity SpCas9 variants, sgRNA expression strategies based on mutant sgRNA scaffold and tRNA processing system, and CRISPR/Cas9-mediated T-DNA integrations. Specificity of CRISPR/Cas9 tools has been a major concern along with the reports of their successful applications. We report unexpected observations of high frequency off-target mutagenesis induced by CRISPR/Cas9 in T1 Arabidopsis mutants although the sgRNA was predicted to have a high specificity score. We also present evidence that the off-target effects were further exacerbated in the T2 progeny. To prevent the off-target effects, we tested and optimized two strategies in Arabidopsis, including introduction of a mCherry cassette for a simple and reliable isolation of Cas9-free mutants and the use of highly specific mutant SpCas9 variants. Optimization of the mCherry vectors and subsequent validation found that fusion of tRNA with the mutant rather than the original sgRNA scaffold significantly improves editing efficiency. We then examined the editing efficiency of eight high-specificity SpCas9 variants in combination with the improved tRNA-sgRNA fusion strategy. Our results suggest that highly specific SpCas9 variants require a higher level of expression than their wild-type counterpart to maintain high editing efficiency. Additionally, we demonstrate that T-DNA can be inserted into the cleavage sites of CRISPR/Cas9 targets with high frequency. Altogether, our results suggest that in plants, continuous attention should be paid to off-target effects induced by CRISPR/Cas9 in current and subsequent generations, and that the tools optimized in this report will be useful in improving genome editing efficiency and specificity in plants and other organisms.
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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.