Abstract
Virus-induced gene silencing (VIGS) is an important tool for the analysis of gene function in plants, which can be adapted for high-throughput functional genomics in model plant species such as Arabidopsis thaliana.
Here we describe the use of the Turnip yellow mosaic virus (TYMV)-derived vector pTY-S that has the ability to induce VIGS in Arabidopsis thaliana. This vector harbors a cDNA copy of the viral genome, in which a unique SnaBI restriction site has been engineered. This site allows the cloning of 80 bp synthetic oligonucleotides corresponding to inverted-repeat fragments of the target gene while retaining the ability of the virus to move systemically. Silencing requires plants to be simply inoculated by abrasion with a few micrograms of intact plasmid DNA, thus precluding the need for in vitro transcription, biolistic, or agroinoculation procedures. This one-step TYMV-based VIGS system is therefore simple to use, cost-effective, and highly consistent, which are important parameters to consider towards the development of high-throughput infection procedures. Another important characteristic of this viral vector is its capacity to infect and induce silencing in meristem tissues.
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Jupin, I. (2013). A Protocol for VIGS in Arabidopsis Thaliana Using a One-Step TYMV-Derived Vector. In: Becker, A. (eds) Virus-Induced Gene Silencing. Methods in Molecular Biology, vol 975. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-278-0_15
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DOI: https://doi.org/10.1007/978-1-62703-278-0_15
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-278-0
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