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Related Concept Videos

RNA Interference01:23

RNA Interference

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RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
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Experimental RNAi02:15

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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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Types of RNA01:20

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Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in regulating gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
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Types of RNA01:23

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Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
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Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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RNAi Interference by dsRNA Injection into Drosophila Embryos
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High-Pressure-Sprayed Double Stranded RNA Does Not Induce RNA Interference of a Reporter Gene.

Veli Vural Uslu1, Alexandra Bassler1, Gabi Krczal1

  • 1AlPlanta-Institute for Plant Research, RLP AgroScience GmbH, Neustadt an der Weinstraße, Germany.

Frontiers in Plant Science
|January 4, 2021
PubMed
Summary

Topical application of naked double-stranded RNA (dsRNA) is a promising pesticide alternative. However, this study found that spraying naked dsRNA onto plants did not effectively trigger RNA interference (RNAi) due to poor cellular uptake.

Keywords:
GFP silencingRNA deliveryRNA interferencedouble stranded RNAsmall RNA sequencing

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Area of Science:

  • Plant molecular biology
  • RNA interference (RNAi) mechanisms
  • Crop protection strategies

Background:

  • RNA interference (RNAi) is a crucial plant defense against pathogens and pests, involving microRNA and small interfering RNA (siRNA) pathways.
  • The siRNA pathway processes double-stranded RNA (dsRNA) into siRNAs, which then activate the RNA-induced silencing complex (RISC) for gene silencing.
  • Artificial activation of RNAi via exogenous dsRNA application offers a potential pesticide substitute, but plant cell wall penetration remains a challenge.

Purpose of the Study:

  • To investigate the effectiveness of topical application of naked dsRNA in triggering RNA interference (RNAi) in plants.
  • To assess the processing of exogenous dsRNA into siRNAs and its uptake by plant cells.
  • To evaluate the potential of dsRNA spraying as a practical, pesticide-free crop protection method.

Main Methods:

  • Utilized a transgenic *Nicotiana benthamiana* line expressing a green fluorescence protein (GFP) reporter gene.
  • Applied naked dsRNA specific to the GFP gene onto plant leaves using high-pressure spraying.
  • Analyzed gene silencing and performed small RNA sequencing (sRNA-seq) to quantify siRNA production and dsRNA processing.

Main Results:

  • High-pressure spraying of naked dsRNA did not induce significant silencing of the GFP reporter gene.
  • Small RNA sequencing revealed that the applied dsRNA was not efficiently processed into siRNAs.
  • Evidence suggests insufficient uptake of dsRNA by plant cells, hindering the initiation of RNAi.

Conclusions:

  • Topical application of naked dsRNA, even with high-pressure spraying, is not an effective method for triggering RNA interference in mature plants.
  • The plant cell wall presents a significant barrier to dsRNA uptake, limiting its potential as a direct foliar pesticide.
  • Further research is needed to develop effective delivery systems for exogenous dsRNA to overcome cellular barriers and enable RNAi-based crop protection.