Enhancement of synergistic gene silencing by RNA interference using branched “3-in-1” trimer siRNA

Nanostructured RNA carrying three different siRNAs was assembled to silence three target genes (Axin, APC, and GSK-3β) in the Wnt/β-catenin signaling pathway. This nanostructured ‘3-in-1’ siRNA showed high activity at a low concentration due to the long-term resistance, and enhancing the effect of RNA interference.

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RNAi-Mediated Management of Whitefly Bemisia tabaci by Oral Delivery of Double-stranded RNAs

Proceedings ◽

10.3390/proceedings2019036011 ◽

2019 ◽

Vol 36 (1) ◽

pp. 11

Author(s):

Jain ◽

Robinson ◽

Mitter

Keyword(s):

Rna Interference ◽

Gene Silencing ◽

Bemisia Tabaci ◽

Oral Delivery ◽

Target Genes ◽

Insect Pests ◽

Control Strategies ◽

Plant Viruses ◽

Potential Game ◽

Viable Approach

The whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) is a significant global pest of economically important vegetable, fibre, and ornamental crops. Whiteflies directly damage the plants by piercing and sucking essential nutrients, indirectly through honeydew secretion and by transmitting more than 200 plant viruses that cause millions of dollars in produce losses per year. Whitefly management is mostly reliant on the heavy use of chemical insecticides. However, this ultimately leads to increasing resistance development, detrimental effects on beneficial insects and biomagnification of ecologically harmful chemicals in the environment. Responding to consumer demands for more selective, less toxic, non-GM insect control strategies, RNA interference (RNAi) has emerged as a potential game-changing solution. The RNA interference (RNAi) is a homology-dependent mechanism of gene silencing that represents a feasible and sustainable technology for the management of insect pests. In the present study, twenty-two whitefly genes were selected based on their essential function in the insect and tested in artificial diet bioassays for mortality and gene silencing efficacy. The nine most effective dsRNA constructs showed moderate-to-high whitefly mortality as compared to negative controls six days post-feeding. qPCR analysis further demonstrated significant knockdown of target gene mRNA expression. Additionally, uptake and spread of fluorescently labelled dsRNA was evident beyond the midgut of the whitefly supporting the systemic spreading of RNAi effectors. Taken together, the oral delivery of dsRNA shows effective RNAi mediated gene silencing of target genes and offers a viable approach for the development of dsRNA biopesticides against hemipteran pest.

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RNA interference as a gene silencing tool to controlTuta absolutain tomato (Solanum lycopersicum)

PeerJ ◽

10.7717/peerj.2673 ◽

2016 ◽

Vol 4 ◽

pp. e2673 ◽

Cited By ~ 21

Author(s):

Roberto A. Camargo ◽

Guilherme O. Barbosa ◽

Isabella Presotto Possignolo ◽

Lazaro E. P. Peres ◽

Eric Lam ◽

...

Keyword(s):

Rna Interference ◽

Gene Silencing ◽

Target Gene ◽

Target Genes ◽

Larval Mortality ◽

Tuta Absoluta ◽

Specific Gene ◽

Gene Transcript ◽

Pest Species ◽

In Planta

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes (Vacuolar ATPase-AandArginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet forT. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliverin planta-transcribed dsRNA to target insect genes.Tuta absolutalarvae that fed on leaves containing dsRNA of the target genes showed an ∼60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage byT. absolutain these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

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RNA interference-mediated gene silencing in murine T cells: in vitro and in vivo validation of proinflammatory target genes

Cell Communication and Signaling ◽

10.1186/1478-811x-6-3 ◽

2008 ◽

Vol 6 (1) ◽

pp. 3 ◽

Cited By ~ 9

Author(s):

Tatjana C Gust ◽

Luisa Neubrandt ◽

Claudia Merz ◽

Khusru Asadullah ◽

Ulrich Zügel ◽

...

Keyword(s):

T Cells ◽

Rna Interference ◽

Gene Silencing ◽

Target Genes ◽

In Vivo Validation

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RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)

10.7287/peerj.preprints.2406 ◽

2016 ◽

Author(s):

Roberto A Camargo ◽

Guilherme O Barbosa ◽

Isabella Presotto Possignolo ◽

Lazaro E. P. Peres ◽

Eric Lam ◽

...

Keyword(s):

Rna Interference ◽

Gene Silencing ◽

Target Gene ◽

Target Genes ◽

Larval Mortality ◽

Tuta Absoluta ◽

Specific Gene ◽

Gene Transcript ◽

Pest Species ◽

In Planta

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes [Vacuolar ATPase-A and Arginine kinase] based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ~60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

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A material-basis study of Aloe vera on the wnt/β-catenin signaling pathway using a knockin/knockout method with high-speed countercurrent chromatography

RSC Advances ◽

10.1039/c7ra06761e ◽

2017 ◽

Vol 7 (62) ◽

pp. 38819-38829 ◽

Cited By ~ 3

Author(s):

Cong Dai ◽

Meng-ping Liu ◽

Wei-jia Zhang ◽

Christopher Wai Kei Lam ◽

Jian-ru Guo ◽

...

Keyword(s):

Signaling Pathway ◽

High Speed ◽

Target Genes ◽

Aloe Vera ◽

Water Extract ◽

Hek293 Cells ◽

Countercurrent Chromatography ◽

Low Concentration ◽

Material Basis

The water extract ofAloe veraat low concentration could activate the wnt/β-catenin signaling pathway and up-regulated the level of active β-catenin protein in hek293 cells, as well as promoting the expression of wnt target genes.

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RNAi-Induced Gene Silencing by Local Electroporation in Targeting Brain Region

Journal of Neurophysiology ◽

10.1152/jn.00161.2004 ◽

2005 ◽

Vol 93 (1) ◽

pp. 594-602 ◽

Cited By ~ 80

Author(s):

Yukio Akaneya ◽

Bin Jiang ◽

Tadaharu Tsumoto

Keyword(s):

Gene Silencing ◽

Brain Region ◽

Target Genes ◽

Short Pulse ◽

Glutamate Release ◽

Long Term Potentiation ◽

Limited Area ◽

Expression Levels ◽

Electrical Pulses

Genetic manipulation for “knockout” (KO) is a useful tool for characterizing a target gene. However, its shortcomings that need to be overcome hinder its easy and ready usage in ordinary laboratories. Here we describe a knockdown technique termed the RNA interference (RNAi)-induced gene silencing by local electroporation (RISLE). Small interfering RNA (siRNA) introduction by electroporation into a specific brain region results in a marked reduction in the expression levels of both the mRNA and protein of the target genes such as GluR2 and Cox-1 without affecting the expression levels of proteins other than that of the target protein or causing pathological changes in the target tissues. The effective electrical pulses are relatively weak, consisting of a strong short pulse and a weak long pulse applied in tandem. RISLE can knock down a gene at the target region, for example, the visual cortex and the CA1 region of the hippocampus, without affecting other regions. Moreover, the knockdown models constructed using this technique have physiological functions consistent with previous findings, that is, glutamate release from presynaptic sites, long-term potentiation (LTP), and long-term depression (LTD). These results suggest that this technique is applicable and characterized by spatial flexibility, temporal accessibility, and ease of establishment of knockdown models. The intactness of the tissue subjected to RISLE is due to the weak electrical pulses applied and the limited area of gene silencing. Thus RISLE may be applicable to disease therapy in the future.

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Faculty Opinions recommendation of Expression of RNA-interference/antisense transgenes by the cognate promoters of target genes is a better gene-silencing strategy to study gene functions in rice.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.9179956.9768058 ◽

2011 ◽

Author(s):

Michael Lassner ◽

Brian McGonigle

Keyword(s):

Rna Interference ◽

Gene Silencing ◽

Target Genes ◽

Gene Functions

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RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)

10.7287/peerj.preprints.2406v1 ◽

2016 ◽

Author(s):

Roberto A Camargo ◽

Guilherme O Barbosa ◽

Isabella Presotto Possignolo ◽

Lazaro E. P. Peres ◽

Eric Lam ◽

...

Keyword(s):

Rna Interference ◽

Gene Silencing ◽

Target Gene ◽

Target Genes ◽

Larval Mortality ◽

Tuta Absoluta ◽

Specific Gene ◽

Gene Transcript ◽

Pest Species ◽

In Planta

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes [Vacuolar ATPase-A and Arginine kinase] based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ~60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

Download Full-text