ArticlePlant physiology2016
An Effective Strategy for Reliably Isolating Heritable and Cas9-Free Arabidopsis Mutants Generated by CRISPR/Cas9-Mediated Genome Editing.
Article in Plant physiology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 144 papers.
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144 citing papers in PubMed.
- CRISPR/Cas9-mediated editing of theGM crops & food · 2026Article
- Transcription factor MdLBD41 negatively regulates resistance to Glomerella leaf spot in apple by suppressing ROS production.The New phytologist · 2026Article
- Systemic and Local Regulation of Root Growth by Vascular Trehalose 6-Phosphate is Correlated With Re-allocation of Primary Metabolites Between Shoots and Roots.Plant, cell & environment · 2026Article
- A simple and reliable method for determining kanamycin sensitivity in Brassica napus.Plant physiology · 2026Article
- Endophytic Flavobacterium promotes root hair development and enhances drought tolerance via an ERF-CEP5 hormonal regulatory module.Nature plants · 2026Article
- Engineered CRISPR-Cas9 Vector for Efficient Agrobacterium-Mediated Transformation of Phanerochaete chrysosporium.Molecular biotechnology · 2026Article
- CRISPR/Cas9-Based Gene Editing in Soybean.Methods in molecular biology (Clifton, N.J.) · 2026Article
- An increase ofeLife · 2025Article
- Callus and endosperm green fluorescence reporter-assisted selection system in maize CRISPR/Cas9 gene editing.BMC plant biology · 2025Article
- Development of an RNA aptamer-assisted CRISPR/Cas9 system for efficiently generating and isolating Cas9-free mutants in plant.PLoS genetics · 2025Article
- Transgene-Killer-CRISPR version 2 (TKC2) eliminates occasional transgene escape by coupling with a RUBY reporter.Plant biotechnology journal · 2025Article
- Article
- CRISPR/Cas system-mediated transgene-free or DNA-free genome editing in plants.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2025Review
- Emerging trends in transgene-free crop development: insights into genome editing and its regulatory overview.Plant molecular biology · 2025Review
- Enhancing drought resilience in crops: mechanistic approaches in the face of climate challenge.Plant molecular biology · 2025Review
- Never-germinating Arabidopsis seeds with LbCas12a-induced mutations in 6 clade A type 2C protein phosphatase genes.Plant physiology · 2025Article
- A major trade-off between growth and defense in Arabidopsis thaliana can vanish in field conditions.PLoS biology · 2025Article
- The metabolite itaconate is a transcriptional and posttranslational modulator of plant metabolism, development, and stress response.Science advances · 2025Article
- Plant genetic transformation: achievements, current status and future prospects.Plant biotechnology journal · 2025Review
- Identification of F-box proteins in ABA- and GA-regulated seed germination: interaction of GASA1 signalling peptide and ABA-induced ubiquitination.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2025Article
84 more citing papers are in PubMed but not listed here.
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5 authors.
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Abstract
Mutations generated by CRISPR/Cas9 in Arabidopsis (Arabidopsis thaliana) are often somatic and are rarely heritable. Isolation of mutations in Cas9-free Arabidopsis plants can ensure the stable transmission of the identified mutations to next generations, but the process is laborious and inefficient. Here, we present a simple visual screen for Cas9-free T2 seeds, allowing us to quickly obtain Cas9-free Arabidopsis mutants in the T2 generation. To demonstrate this in principle, we targeted two sites in the AUXIN-BINDING PROTEIN1 (ABP1) gene, whose function as a membrane-associated auxin receptor has been challenged recently. We obtained many T1 plants with detectable mutations near the target sites, but only a small fraction of T1 plants yielded Cas9-free abp1 mutations in the T2 generation. Moreover, the mutations did not segregate in Mendelian fashion in the T2 generation. However, mutations identified in the Cas9-free T2 plants were stably transmitted to the T3 generation following Mendelian genetics. To further simplify the screening procedure, we simultaneously targeted two sites in ABP1 to generate large deletions, which can be easily identified by PCR. We successfully generated two abp1 alleles that contained 1,141- and 711-bp deletions in the ABP1 gene. All of the Cas9-free abp1 alleles we generated were stable and heritable. The method described here allows for effectively isolating Cas9-free heritable CRISPR mutants in Arabidopsis.
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