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Related Concept Videos

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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Double-stranded RNA Oral Delivery Methods to Induce RNA Interference in Phloem and Plant-sap-feeding Hemipteran Insects
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Optimal dsRNA Concentration for RNA Interference in Asian Citrus Psyllid.

Esmaeil Saberi1, Mosharrof Mondal2,3, Jorge R Paredes-Montero4,5

  • 1Southwest Florida Research and Education Center, Department of Entomology and Nematology, IFAS, University of Florida, Immokalee, FL 34142, USA.

Insects
|January 22, 2024
PubMed
Summary

Double-stranded RNA (dsRNA) biopesticides show promise for controlling the Asian citrus psyllid (ACP), a vector for citrus greening disease. This study determined the minimal effective dsRNA concentration for significant gene knockdown in ACP, identifying 200 ng/µL as optimal for RNA interference (RNAi) penetrance.

Keywords:
Diaphorina citriRNA interferencedsRNA biopesticidegene knockdownpest control

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Area of Science:

  • Entomology
  • Plant Pathology
  • Molecular Biology

Background:

  • The Asian citrus psyllid (ACP) transmits *Candidatus* Liberibacter asiaticus, causing citrus greening disease.
  • Double-stranded RNA (dsRNA) biopesticides offer an RNA interference (RNAi) mechanism as an alternative to conventional insecticides.
  • Standardized dsRNA screening requires determining minimal effective concentrations for RNAi penetrance and measurable phenotypes like gene knockdown.

Purpose of the Study:

  • To evaluate gene knockdown in ACP following exposure to varying dsRNA concentrations.
  • To identify the minimal effective dsRNA concentration for optimal RNAi penetrance in ACP.

Main Methods:

  • ACP adults and third instar nymphs were fed dsRNA targeting clathrin heavy chain (CHC), vacuolar ATPase subunit A (vATPase-A), and sucrose non-fermenting protein 7 (Snf7) genes.
  • Ingestion-access periods (IAP) of 48 hours were followed by 5 days on untreated host plants.
  • Gene knockdown was quantified using qPCR across dsRNA concentrations ranging from 10 to 500 ng/µL.

Main Results:

  • Significant gene knockdown (p < 0.05) was observed in third instar nymphs (12-34%) and adults (18-39%) at dsRNA concentrations of 10-500 ng/µL and 100-500 ng/µL, respectively.
  • The threshold concentration for significant gene knockdown and adult mortality was determined to be 200 ng/µL.
  • This concentration (200 ng/µL) was identified as indicative of optimal RNAi penetrance in ACP.

Conclusions:

  • dsRNA biopesticides can effectively induce gene knockdown in ACP, suggesting a viable RNAi-based pest control strategy.
  • The optimal dsRNA concentration for effective RNAi penetrance in ACP is approximately 200 ng/µL.
  • Further research can utilize these findings to develop targeted dsRNA biopesticides against ACP and citrus greening disease.