ArticlePlant & cell physiology2017
In Planta Processing of the SpCas9-gRNA Complex.
Article in Plant & cell physiology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 49 citations in OpenAlex.
- Efficient CRISPR-Cas9 delivery and transgene-free multiplex genome editing in plants using cymbidium mosaic virus-derived vectors.The Plant journal : for cell and molecular biology · 2026Article
- Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding.Plants (Basel, Switzerland) · 2023Review
- Building Blocks of Artificial CRISPR-Based Systems beyond Nucleases.International journal of molecular sciences · 2022Review
- Functional Allele Validation by Gene Editing to Leverage the Wealth of Genetic Resources for Crop Improvement.International journal of molecular sciences · 2022Review
- Multiplex knockout of trichome-regulating MYB duplicates in hybrid poplar using a single gRNA.Plant physiology · 2022Article
- Two efficient CRISPR/Cas9 systems for gene editing in soybean.Transgenic research · 2021Article
- A bacterial small RNA regulates the adaptation of Helicobacter pylori to the host environment.Nature communications · 2021Article
- Multiplexed Genome EditingFrontiers in plant science · 2021Article
- Potato Virus X Vector-Mediated DNA-Free Genome Editing in Plants.Plant & cell physiology · 2020Article
- Applications of CRISPR-Cas in agriculture and plant biotechnology.Nature reviews. Molecular cell biology · 2020Review
- Excising the Mystery of Single-Guide RNA Processing.Plant physiology · 2020Article
- Native Processing of Single Guide RNA Transcripts to Create Catalytic Cas9/Single Guide RNA Complexes in Planta.Plant physiology · 2020Article
- Review
- Intron-Based Single Transcript Unit CRISPR Systems for Plant Genome Editing.Rice (New York, N.Y.) · 2020Article
- A CRISPR/dCas9 toolkit for functional analysis of maize genes.Plant methods · 2020Article
- An update and perspectives on the use of promoters in plant genetic engineering.Journal of biosciences · 2020Review
- Enhanced FnCas12a-Mediated Targeted Mutagenesis Using crRNA With Altered Target Length in Rice.Frontiers in genome editing · 2020Article
- Strategies to Increase On-Target and Reduce Off-Target Effects of the CRISPR/Cas9 System in Plants.International journal of molecular sciences · 2019Review
- Single transcript unit CRISPR 2.0 systems for robust Cas9 and Cas12a mediated plant genome editing.Plant biotechnology journal · 2019Article
- Gene editing in plants: progress and challenges.National science review · 2019Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9)-mediated genome editing in plants, Streptococcus pyogenes Cas9 (SpCas9) protein and the required guide RNA (gRNA) are, in most cases, expressed from a stably integrated transgene. Generally, SpCas9 protein is expressed from an RNA polymerase (pol) II promoter, while gRNA is expressed from a pol III promoter. However, pol III promoters have not been much characterized other than in model plants, making it difficult to select appropriate promoters for specific applications, while pol II transcripts have to be processed to generate functional gRNAs. Recently, successful processing of a pol II transcript into functional gRNAs using ribozyme or Csy4-RNA cleavage systems has been demonstrated. Here, we show that functional gRNAs can be efficiently processed using SpCas9 protein and plant endogenous RNA cleavage systems without the need for a specific RNA processing system. In our system, SpCas9 RNA and gRNA are both transcribed as a single RNA using a single pol II promoter; translated SpCas9 protein can be bound to this RNA and, finally, extra RNA sequences are trimmed by plant RNA processing systems to form a functional SpCas9-gRNA complex. The efficiency of targeted mutagenesis using our novel SpCas9-gRNA fused system was comparable with that of the SpCas9-gRNA system with ribozyme sequence, achieving rates of up to 100% in rice. Our results could be useful in developing stable SpCas9-gRNA expression systems and in RNA virus vector-mediated genome editing systems in plants.
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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.