ArticlePlant physiology2010
Rapid assessment of gene function in the circadian clock using artificial microRNA in Arabidopsis mesophyll protoplasts.
Article in Plant physiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers.
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Who cites it
55 citing papers in PubMed, 104 citations in OpenAlex.
- Enhanced microRNA accumulation and gene silencing efficiency through optimized precursor base pairing.The Plant journal : for cell and molecular biology · 2026Article
- A CRISPR/Cas9-induced restoration of bioluminescence reporter system for single-cell gene expression analysis in plants.Scientific reports · 2025Article
- TOC1 phosphorylation disproportionally enhances chromatin binding at rhythmic gene promoters.Science advances · 2025Article
- Molecular mechanisms underlying gene regulatory variation of maize metabolic traits.The Plant cell · 2024Article
- Comprehensive transcriptomic analysis of age-, dark-, and salt-induced senescence reveals underlying mechanisms and key regulators of leaf senescence inFrontiers in plant science · 2023Article
- The NaProceedings of the National Academy of Sciences of the United States of America · 2022Article
- Modeling temporal and hormonal regulation of plant transcriptional response to wounding.The Plant cell · 2022Article
- Establishing an efficient protoplast transient expression system for investigation of floral thermogenesis in aroids.Plant cell reports · 2022Article
- Rapid Investigation of Functional Roles of Genes in Regulation of Leaf Senescence Using Arabidopsis Protoplasts.Frontiers in plant science · 2022Article
- Agrobacterium-Mediated Seedling Transformation to Measure Circadian Rhythms in Arabidopsis.Methods in molecular biology (Clifton, N.J.) · 2022Article
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- The TSN1 Binding Protein RH31 Is a Component of Stress Granules and Participates in Regulation of Salt-Stress Tolerance inFrontiers in plant science · 2021Article
- KHZ1 and KHZ2, novel members of the autonomous pathway, repress the splicing efficiency of FLC pre-mRNA in Arabidopsis.Journal of experimental botany · 2020Article
- Attenuated TOR signaling lengthens circadian period inPlant signaling & behavior · 2020Article
- Casein kinase 1 family regulates PRR5 and TOC1 in the Arabidopsis circadian clock.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- Basic LEUCINE ZIPPER TRANSCRIPTION FACTOR67 TransactivatesThe Plant cell · 2019Article
- Genome-wide analysis of HSP70 family genes in cabbage (Brassica oleracea var. capitata) reveals their involvement in floral development.BMC genomics · 2019Article
- The Plant Circadian Oscillator.Biology · 2019Review
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Corrections and comments
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Authors and funding
2 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Rapid assessment of the effect of reduced levels of gene products is often a bottleneck in determining how to proceed with an interesting gene candidate. Additionally, gene families with closely related members can confound determination of the role of even a single one of the group. We describe here an in vivo method to rapidly determine gene function using transient expression of artificial microRNAs (amiRNAs) in Arabidopsis (Arabidopsis thaliana) mesophyll protoplasts. We use a luciferase-based reporter of circadian clock activity to optimize and validate this system. Protoplasts transiently cotransfected with promoter-luciferase and gene-specific amiRNA plasmids sustain free-running rhythms of bioluminescence for more than 6 d. Using both amiRNA plasmids available through the Arabidopsis Biological Resource Center, as well as custom design of constructs using the Weigel amiRNA design algorithm, we show that transient knockdown of known clock genes recapitulates the same circadian phenotypes reported in the literature for loss-of-function mutant plants. We additionally show that amiRNA designed to knock down expression of the casein kinase II β-subunit gene family lengthens period, consistent with previous reports of a short period in casein kinase II β-subunit overexpressors. Our results demonstrate that this system can facilitate a much more rapid analysis of gene function by obviating the need to initially establish stably transformed transgenics to assess the phenotype of gene knockdowns. This approach will be useful in a wide range of plant disciplines when an endogenous cell-based phenotype is observable or can be devised, as done here using a luciferase reporter.
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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.