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Experimental RNAi02:15

Experimental RNAi

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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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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.
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Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
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Double-stranded RNA Oral Delivery Methods to Induce RNA Interference in Phloem and Plant-sap-feeding Hemipteran Insects
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Why is oral-induced RNAi inefficient in Diatraea saccharalis? A possible role for DsREase and other nucleases.

Manoely Abreu Reis1, Daniel David Noriega2, Gessica Dos Santos Alves1

  • 1Departamento de Agronomia-Entomologia, Universidade Federal Rural de Pernambuco - UFRPE, Recife, Brazil.

Pesticide Biochemistry and Physiology
|August 16, 2022
PubMed
Summary
This summary is machine-generated.

RNA interference (RNAi) is inefficient in the sugarcane borer, Diatraea saccharalis, due to rapid degradation of double-stranded RNA (dsRNA) by gut nucleases. This study identifies specific nucleases contributing to RNAi failure in this pest.

Keywords:
NucleaseRNA interferenceRNAi efficiencySugarcane borerdsRNA stability

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Area of Science:

  • Entomology
  • Molecular Biology
  • Pest Management

Background:

  • RNAi efficiency varies in insects, especially lepidopterans.
  • Degradation of double-stranded RNA (dsRNA) by nucleases limits RNAi efficacy.
  • Effective gene silencing via oral dsRNA delivery is poorly understood in Diatraea saccharalis.

Purpose of the Study:

  • To investigate the feasibility of gene silencing through dsRNA feeding in D. saccharalis.
  • To identify factors limiting RNAi efficiency in this species.
  • To understand the role of gut nucleases in dsRNA degradation.

Main Methods:

  • Oral delivery of dsRNA targeting DsJHE and DsCHS1 genes.
  • Analysis of dsRNA degradation in gut juice.
  • Identification and expression analysis of potential nucleases (DsNucleases, DsREase).

Main Results:

  • Discrete gene knockdown only observed for DsCHS1 with high dsRNA doses and prolonged exposure.
  • dsRNA was rapidly degraded in D. saccharalis gut juice.
  • Four nucleases were identified, with DsNuclease1 and DsREase highly expressed in the larval gut.
  • DsREase expression increased with artificial diet or dsRNA injection.

Conclusions:

  • Gut nuclease activity is a significant barrier to RNAi efficacy in D. saccharalis.
  • Specific nucleases, particularly DsREase, contribute to dsRNA degradation.
  • Overcoming nuclease activity is crucial for developing RNAi-based pest control strategies in lepidopterans.