ArticleFrontiers in plant science2016
COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants.
Article in Frontiers in plant science, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- Robust quantification of multiplexed fluorescent protein-based biosensors in plant tissues.The Plant journal : for cell and molecular biology · 2026Article
- A mini-TGA protein modulates gene expression through heterogeneous association with transcription factors.Plant physiology · 2023Article
- A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants.Plant communications · 2023Article
- Heterologous mogrosides biosynthesis in cucumber and tomato by genetic manipulation.Communications biology · 2023Article
- Pyramiding stacking of multigenes (PSM): a simple, flexible and efficient multigene stacking system based on Gibson assembly and gateway cloning.Frontiers in bioengineering and biotechnology · 2023Article
- A New Set of Golden-Gate-Based Organelle Marker Plasmids for Colocalization Studies in Plants.Plants (Basel, Switzerland) · 2022Article
- EXTRA LARGE G-PROTEIN2 mediates cell death and hyperimmunity in the chitin elicitor receptor kinase 1-4 mutant.Plant physiology · 2022Article
- Compendium on Food Crop Plants as a Platform for Pharmaceutical Protein Production.International journal of molecular sciences · 2022Review
- An efficient papaya leaf distortion mosaic potyvirus vector for virus-induced gene silencing in papaya.Horticulture research · 2021Article
- Live-cell RESOLFT nanoscopy of transgenicPlant direct · 2020Article
- Recombinase-mediated integration of a multigene cassette in rice leads to stable expression and inheritance of the stacked locus.Plant direct · 2020Article
- Plant Synthetic Metabolic Engineering for Enhancing Crop Nutritional Quality.Plant communications · 2020Review
- Quantitative Hormone Signaling Output Analyses ofFrontiers in plant science · 2020Article
- Nimble Cloning: A Simple, Versatile, and Efficient System for Standardized Molecular Cloning.Frontiers in bioengineering and biotechnology · 2019Article
- I see the light! Fluorescent proteins suitable for cell wall/apoplast targeting inPlant direct · 2019Article
- Plant X-tender: An extension of the AssemblX system for the assembly and expression of multigene constructs in plants.PloS one · 2018Article
- Article
- A Tightly Regulated Genetic Selection System with Signaling-Active Alleles of Phytochrome B.Plant physiology · 2017Article
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3 authors.
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No grant is acknowledged in the PubMed record.
Abstract
Advancing basic and applied plant research requires the continuous innovative development of the available technology toolbox. Essential components of this toolbox are methods that simplify the assembly, delivery, and expression of multiple transgenes of interest. To allow simultaneous and directional multigene assembly on the same plant transformation vector, several strategies based on overlapping sequences or restriction enzymes have recently been developed. However, the assembly of homologous and repetitive DNA sequences can be inefficient and the frequent occurrence of target sequences recognized by commonly used restriction enzymes can be a limiting factor. Here, we noted that recognition sites for the restriction enzyme SfiI are rarely occurring in plant genomes. This fact was exploited to establish a multigene assembly system called "COLORFUL-Circuit." To this end, we developed a set of binary vectors which provide a flexible and cost efficient cloning platform. The gene expression cassettes in our system are flanked with unique SfiI sites, which allow simultaneous multi-gene cassette assembly in a hosting binary vector. We used COLORFUL-Circuit to transiently and stably express up to four fluorescent organelle markers in addition to a selectable marker and analyzed the impact of assembly design on coexpression efficiency. Finally, we demonstrate the utility of our optimized "COLORFUL-Circuit" system in an exemplary case study, in which we monitored simultaneously the subcellular behavior of multiple organelles in a biotrophic plant-microbe interaction by Confocal Laser Scanning Microscopy.
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