ArticlePloS one2014
A modular plasmid assembly kit for multigene expression, gene silencing and silencing rescue in plants.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 77 papers.
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77 citing papers in PubMed.
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- Mapping genetic modifiers of epimutation rates identifies VIM2/4 as dosage-sensitive negative regulators of CG methylation maintenance.Nature plants · 2026Article
- The Arabidopsis SPFH protein HIR2 modulates receptor signaling and plasma membrane organization.The EMBO journal · 2026Article
- The plant Golden Gate toolkit: compatibility, interoperability, and harmonized assembly in plant synthetic biology.Plant methods · 2026Review
- dSaCas9 enables enhanced transcriptional activation in Nicotiana benthamiana compared to its dSpCas9 ortholog.BMC plant biology · 2026Article
- Cross-kingdom RNA interference promotes arbuscular mycorrhiza development.Nature plants · 2026Article
- A novel cis-element enabled bacterial uptake by plant cells.Nature plants · 2026Article
- Engineering and comparison of cas12a-based genome editing systems in plants.The Plant journal : for cell and molecular biology · 2025Article
- Reverse engineering of the pattern recognition receptor FLS2 reveals key design principles of broader recognition spectra against evading flg22 epitopes.Nature plants · 2025Article
- The transcription factor WRKY25 can act as redox switch to drive the expression of WRKY53 during leaf senescence in arabidopsis.Scientific reports · 2025Article
- Article
- Genome modification in plant mitochondria.Plant physiology · 2025Review
- Medicago Mting1 Mting2 double knockout mutants are extremely dwarfed and never flower implicating essential MtING functions in growth and flowering.BMC plant biology · 2025Article
- Ethylene promotes SMAX1 accumulation to inhibit arbuscular mycorrhiza symbiosis.Nature communications · 2025Article
- A small signaling domain controls PPIP5K phosphatase activity in phosphate homeostasis.Nature communications · 2025Article
- Optimized LC-MS method for simultaneous polyamine profiling and ADC/ODC activity quantification and evidence that ADCs are indispensable for flower development in tomato.Frontiers in plant science · 2025Article
- A Simple RUBY Reporter System in Nicotiana benthamiana for Studying Inositol Pyrophosphate Dynamics.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Assembling DNA Plasmids with the Multi-Kingdom (MK) Cloning System.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Golden Gate Cloning of Multigene Constructs Using the Modular Cloning System MoClo.Methods in molecular biology (Clifton, N.J.) · 2025Article
17 more citing papers are in PubMed but not listed here.
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Golden Gate (GG) modular assembly approach offers a standardized, inexpensive and reliable way to ligate multiple DNA fragments in a pre-defined order in a single-tube reaction. We developed a GG based toolkit for the flexible construction of binary plasmids for transgene expression in plants. Starting from a common set of modules, such as promoters, protein tags and transcribed regions of interest, synthetic genes are assembled, which can be further combined to multigene constructs. As an example, we created T-DNA constructs encoding multiple fluorescent proteins targeted to distinct cellular compartments (nucleus, cytosol, plastids) and demonstrated simultaneous expression of all genes in Nicotiana benthamiana, Lotus japonicus and Arabidopsis thaliana. We assembled an RNA interference (RNAi) module for the construction of intron-spliced hairpin RNA constructs and demonstrated silencing of GFP in N. benthamiana. By combination of the silencing construct together with a codon adapted rescue construct into one vector, our system facilitates genetic complementation and thus confirmation of the causative gene responsible for a given RNAi phenotype. As proof of principle, we silenced a destabilized GFP gene (dGFP) and restored GFP fluorescence by expression of a recoded version of dGFP, which was not targeted by the silencing construct.
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