Artificial trans-acting small interfering RNA: a tool for plant biology study and crop improvements

Zhang, Zhanyuan J.

doi:10.1007/s00425-014-2054-x

Cited by 38 publications

(22 citation statements)

References 47 publications

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“…amiRNAs were first used to confer antiviral resistance in transgenic plants (Niu et al ., ); however, virus sequence variants accumulating mutations in the amiRNA target site (TS) may overcome this resistance (Simon‐Mateo and Garcia, ; Lin et al ., ; Lafforgue et al ., ). More recently, syn‐tasiRNAs have emerged as a promising antiviral tool because of their unique multiplexing capability (Zhang, ; Carbonell, 2019b) as well as for the availability of high‐throughput cloning strategies and automated design tools for the simple generation of syn‐tasiRNA constructs (Carbonell et al ., ; Carbonell, 2019a). The simultaneous co‐expression from a single precursor of several syn‐tasiRNAs capable of targeting multiple sites within a single viral RNA or within different viral RNAs should induce more effective, durable, and broad antiviral resistance (Carbonell et al ., ).…”

Section: Introductionmentioning

confidence: 99%

Multi‐targeting of viral RNAs with synthetic trans‐acting small interfering RNAs enhances plant antiviral resistance

2019

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Summary RNA interference (RNAi)‐based tools are used in multiple organisms to induce antiviral resistance through the sequence‐specific degradation of target RNAs by complementary small RNAs. In plants, highly specific antiviral RNAi‐based tools include artificial microRNAs (amiRNAs) and synthetic trans‐acting small interfering RNAs (syn‐tasiRNAs). syn‐tasiRNAs have emerged as a promising antiviral tool allowing for the multi‐targeting of viral RNAs through the simultaneous expression of several syn‐tasiRNAs from a single precursor. Here, we compared in tomato plants the effects of an amiRNA construct expressing a single amiRNA and a syn‐tasiRNA construct expressing four different syn‐tasiRNAs against Tomato spotted wilt virus (TSWV), an economically important pathogen affecting tomato crops worldwide. Most of the syn‐tasiRNA lines were resistant to TSWV, whereas the majority of the amiRNA lines were susceptible and accumulated viral progenies with mutations in the amiRNA target site. Only the two amiRNA lines with higher amiRNA accumulation were resistant, whereas resistance in syn‐tasiRNA lines was not exclusive of lines with high syn‐tasiRNA accumulation. Collectively, these results suggest that syn‐tasiRNAs induce enhanced antiviral resistance because of the combined silencing effect of each individual syn‐tasiRNA, which minimizes the possibility that the virus simultaneously mutates all different target sites to fully escape each syn‐tasiRNA.

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Section: Introductionmentioning

confidence: 99%

Multi‐targeting of viral RNAs with synthetic trans‐acting small interfering RNAs enhances plant antiviral resistance

2019

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“…siRNA is thought to be an RNAbased immune mechanism that fights off nucleic acid invaders and maintains genome stability. 12 In our study, two recombinant adenoviral vectors based on the GV120 vector ( Figure 1a; purchased from GeneChem Co, Ltd, Shanghai, China) were constructed to silence the mouse S100A4 gene (Ad-S100A4-RNAi) and to express the green fluorescence protein gene (Ad-GFP; negative control). The target sequence is 5′-TGA GCA ACT TGG ACA GCA A-3′, and the negative control sequence is 5′-TTC TCC GAA CGT GTC ACG T-3′.…”

Section: Recombinant Adenoviral Vector Constructionmentioning

confidence: 99%

Silencing of S100A4, a metastasis-associated protein, inhibits retinal neovascularization via the downregulation of BDNF in oxygen-induced ischaemic retinopathy

Cheng

Xing

2016

Eye

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Background To investigate the underlying mechanism of S100A4 function and whether it has a role in retinal neovascularization (RNV) in a mouse model of oxygen-induced retinopathy (OIR). Methods Retinas from a mouse model of OIR were treated with and without an intravitreous injection of adenoviral-S100A4-RNAi or adenoviral green fluorescence protein (GFP) at postnatal day 12 (P12). At P17, the efficacy of adenoviral gene transfer was assessed using fluorescence microscopy and western blot analysis. RNV was evaluated by wholemount immunofluorescence staining of the mouse retina and by counting the number of pre-retinal neovascular cells. Protein and mRNA expression levels of S100A4, brainderived growth factor (BDNF), and vascular endothelial growth factor (VEGF) were measured using western blot analysis and real-time PCR. Results Retinal S100A4 levels were positively correlated with the progression of RNV. In the OIR-S100A4-RNAi group, both protein and mRNA expression levels of S100A4 in the retina significantly decreased at P17 compared with those in the OIR group. Ad-S100A4-RNAi transfer was clearly demonstrated by GFP fluorescence in many layers of the retina 5 days after the Ad-S100A4-RNAi transfer. Whole-mount immunofluorescence staining of the retina and quantification of the pre-retinal neovascular cells demonstrated that RNV was significantly inhibited. Meanwhile, the levels of the transcription and translation of BDNF,

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“…Although amiRNAs derive from DICER‐LIKE1 (DCL1) cleavage of miRNA precursors with foldback structures, syn‐tasiRNAs are produced in a multi‐step process including: (i) cleavage of a TAS precursor by an miRNA/ARGONAUTE (AGO) complex, (ii) conversion of one of the cleavage products to double‐stranded RNA (dsRNA) by RNA‐DEPENDENT RNA POLYMERASE 6, and (iii) sequential processing by DCL4 of the dsRNA into 21‐nucleotide phased syn‐tasiRNAs in register with the miRNA‐guided cleavage site. One of the strands of the amiRNA or syn‐tasiRNA duplex associates with an AGO protein to silence highly sequence complementary transcripts, usually through AGO‐mediated endonucleolytic cleavage (Tiwari et al ., ; Zhang, ).…”

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confidence: 99%

“…register with the miRNA-guided cleavage site. One of the strands of the amiRNA or syn-tasiRNA duplex associates with an AGO protein to silence highly sequence complementary transcripts, usually through AGO-mediated endonucleolytic cleavage (Tiwari et al, 2014;Zhang, 2014).…”

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confidence: 99%

Artificial microRNAs and synthetic trans‐acting small interfering RNAs interfere with viroid infection

Carbonell

Daròs

2017

Molecular Plant Pathology

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SUMMARYArtificial microRNAs (amiRNAs) and synthetic trans-acting small interfering RNAs (syn-tasiRNAs) are two classes of artificial small RNAs (sRNAs) engineered to silence endogenous transcripts as well as viral RNAs in plants. Here, we explore the possibility of using amiRNAs and syn-tasiRNAs to specifically interfere with infections by viroids, small (250-400-nucleotide) non-coding circular RNAs with compact secondary structure infecting a wide range of plant species. The combined use of recent highthroughput methods for artificial sRNA construct generation and the Potato spindle tuber viroid (PSTVd)-Nicotiana benthamiana pathosystem allowed for the simple and time-effective screening of multiple artificial sRNAs targeting sites distributed along PSTVd RNAs of (1) or (-) polarity. The majority of amiRNAs were highly active in agroinfiltrated leaves when co-expressed with an infectious PSTVd transcript, as were syn-tasiRNAs derived from a construct including the five most effective amiRNA sequences. A comparative analysis showed that the effects of the most effective amiRNA and of the syn-tasiRNAs were similar in agroinfiltrated leaves, as well as in upper non-agroinfiltrated leaves in which PSTVd accumulation was significantly delayed. These results suggest that amiRNAs and syntasiRNAs can be used effectively to control viroid infections in plants.Keywords: artificial microRNA, pathogen resistance, PSTVd, RNA silencing, small RNA, synthetic trans-acting siRNA, viroid.Artificial microRNAs (amiRNAs) and synthetic trans-acting small interfering RNAs (syn-tasiRNAs) are two classes of artificial small RNAs (sRNAs) engineered to silence specific transcripts in plants. They are produced in planta by the expression of a functional miRNA or tasiRNA precursor with modified miRNA/miRNA* or tasiRNA sequences, respectively. amiRNAs and syn-tasiRNAs are functionally similar, but are generated differently. Although amiRNAs derive from DICER-LIKE1 (DCL1) cleavage of miRNA precursors with foldback structures, syn-tasiRNAs are produced in a

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Artificial trans-acting small interfering RNA: a tool for plant biology study and crop improvements

Cited by 38 publications

References 47 publications

Multi‐targeting of viral RNAs with synthetic trans‐acting small interfering RNAs enhances plant antiviral resistance

Multi‐targeting of viral RNAs with synthetic trans‐acting small interfering RNAs enhances plant antiviral resistance

Silencing of S100A4, a metastasis-associated protein, inhibits retinal neovascularization via the downregulation of BDNF in oxygen-induced ischaemic retinopathy

Artificial microRNAs and synthetic trans‐acting small interfering RNAs interfere with viroid infection

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