ArticlePlant methods2016
A simple test for the cleavage activity of customized endonucleases in plants.
Article in Plant methods, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 45 citations in OpenAlex.
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- Targeted genome modification in protoplasts of a tea cultivar Kolkhida using RNA-guided Cas9 endonuclease.AoB PLANTS · 2025Article
- A simple and efficient TALEN system for genome editing in plants.Plant molecular biology · 2025Article
- Accumulation of Anthocyanin in the Aleurone of Barley Grains by Targeted Restoration of theInternational journal of molecular sciences · 2024Article
- HOMEOBOX2, the paralog of SIX-ROWED SPIKE1/HOMEOBOX1, is dispensable for barley spikelet development.Journal of experimental botany · 2024Article
- Anatomical insights into the vascular layout of the barley rachis: implications for transport and spikelet connection.Annals of botany · 2024Article
- Ribonucleoprotein (RNP)-Mediated Allele Replacement in Barley (Hordeum vulgare L.) Leaves.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Hairy CRISPR: Genome Editing in Plants Using Hairy Root Transformation.Plants (Basel, Switzerland) · 2021Review
- Targeted genome modifications in cereal crops.Breeding science · 2021Article
- Functional Validation ofInternational journal of molecular sciences · 2021Article
- An improved biolistic delivery and analysis method for evaluation of DNA and CRISPR-Cas delivery efficacy in plant tissue.Scientific reports · 2021Article
- Transcriptional landscapes of floral meristems in barley.Science advances · 2021Article
- CRISPR/Cas9-Induced Loss-of-Function Mutation in the BarleyFrontiers in plant science · 2021Article
- Mutation of theFrontiers in plant science · 2021Article
- Site-directed mutagenesis in bread and durum wheat via pollination by cas9/guide RNA-transgenic maize used as haploidy inducer.Plant biotechnology journal · 2020Article
- Conversion of hulled into naked barley by Cas endonuclease-mediated knockout of the NUD gene.BMC plant biology · 2020Article
- Biolistic Approach for Transient Gene Expression Studies in Plants.Methods in molecular biology (Clifton, N.J.) · 2020Review
- Leaf Variegation and Impaired Chloroplast Development Caused by a Truncated CCT Domain Gene inThe Plant cell · 2019Article
- Cas Endonuclease Technology-A Quantum Leap in the Advancement of Barley and Wheat Genetic Engineering.International journal of molecular sciences · 2019Review
- Modification of Barley Plant Productivity Through Regulation of Cytokinin Content by Reverse-Genetics Approaches.Frontiers in plant science · 2018Article
Corrections and comments
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Authors and funding
12 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundAlthough customized endonucleases [transcription activator-like effector nucleases (TALENs) and RNA-guided endonucleases (RGENs)] are known to be effective agents of mutagenesis in various host plants, newly designed endonuclease constructs require some pre-validation with respect to functionality before investing in the creation of stable transgenic plants.
resultsA simple, biolistics-based leaf epidermis transient expression test has been developed, based on reconstituting the translational reading frame of a mutated, non-functional yfp reporter gene. Quantification of mutation efficacy was made possible by co-bombarding the explant with a constitutive mCherry expression cassette, thereby allowing the ratio between the number of red and yellow fluorescing cells to serve as a metric for mutation efficiency. Challenging either stable mutant alleles of a compromised version of gfp in tobacco and barley or the barley MLO gene with TALENs/RGENs confirmed the capacity to induce site-directed mutations.
conclusionsA convenient procedure to assay the cleavage activity of customized endonucleases has been established. The system is independent of the endonuclease platform and operates in both di- and monocotyledonous hosts. It not only enables the validation of a TALEN/RGEN's functionality prior to the creation of stable mutants, but also serves as a suitable tool to optimize the design of endonuclease constructs.
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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.