ArticleMethods in molecular biology (Clifton, N.J.)2019
Fluorescence Marker-Assisted Isolation of Cas9-Free and CRISPR-Edited Arabidopsis Plants.
Article in Methods in molecular biology (Clifton, N.J.), 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- CRISPR/Cas9-mediated editing of theGM crops & food · 2026Article
- Development of an RNA aptamer-assisted CRISPR/Cas9 system for efficiently generating and isolating Cas9-free mutants in plant.PLoS genetics · 2025Article
- CRISPR/Cas system-mediated transgene-free or DNA-free genome editing in plants.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2025Review
- Time series single-cell transcriptional atlases reveal cell fate differentiation driven by light in Arabidopsis seedlings.Nature plants · 2023Article
- Novel Plant Breeding Techniques Shake Hands with Cereals to Increase Production.Plants (Basel, Switzerland) · 2022Review
- Advances in gene editing without residual transgenes in plants.Plant physiology · 2022Article
- Advances in application of genome editing in tomato and recent development of genome editing technology.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2021Review
- CRISPR-Based Genome Editing Tools: Insights into Technological Breakthroughs and Future Challenges.Genes · 2021Review
- Application and future perspective of CRISPR/Cas9 genome editing in fruit crops.Journal of integrative plant biology · 2020Review
- Repurposing of Anthocyanin Biosynthesis for Plant Transformation and Genome Editing.Frontiers in genome editing · 2020Article
- Technological breakthroughs in generating transgene-free and genetically stable CRISPR-edited plants.aBIOTECH · 2020Review
- Article
- Using Genetic Engineering Techniques to Develop Banana Cultivars With Fusarium Wilt Resistance and Ideal Plant Architecture.Frontiers in plant science · 2020Review
- Improving Nutritional and Functional Quality by Genome Editing of Crops: Status and Perspectives.Frontiers in plant science · 2020Review
- Identification of Transgene-Free CRISPR-Edited Plants of Rice, Tomato, andFrontiers in plant science · 2019Article
Corrections and comments
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Authors and funding
2 authors.
Funding
Abstract
CRISPR/Cas9 gene editing technology has successfully introduced modifications at target DNA sequences in many plant species including Arabidopsis. After the target gene is edited, the CRISPR/Cas9 construct needs to be removed to ensure genetic stability and to gain any regulatory approval for commercial applications. However, removal of the transgenes by genetic segregation, backcross, and genotyping is very laborious and time-consuming. The methods we report here allow fast and effective isolation of transgene-free T2 Arabidopsis plants with the desired modifications at the target genes. We express a fluorescence protein mCherry under the control of a seed-specific promoter At2S3 and placed the cassette into the CRISPR/Cas9 vector. Therefore, we can use mCherry as a proxy for the presence of Cas9, and we are able to visually isolate the Cas9-free Arabidopsis plants with heritable mutations at the T2 generation. We targeted two sites in the ABP1 gene to demonstrate the effectiveness of our approach.
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