Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Xin Zhang; Keshava Mysore; Ellen Flannery; Kristin Michel; David W. Severson; Kun Yan Zhu; Molly Duman-Scheel

doi:10.3791/52523

Chitosane / ARN interférant nanoparticules Mediated Gene Silencing dans la maladie vectorielle la...

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Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Chitosane / ARN interférant nanoparticules Mediated Gene Silencing dans la maladie vectorielle larves de moustiques

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08:36 min

March 25, 2015

DOI: 10.3791/52523-v

Xin Zhang*¹, Keshava Mysore*^2,3, Ellen Flannery^3,4, Kristin Michel¹, David W. Severson^3,4, Kun Yan Zhu⁵, Molly Duman-Scheel^2,3,4

¹Division of Biology,Kansas State University, ²Department of Medical and Molecular Genetics,Indiana University School of Medicine, ³Eck Institute for Global Health,University of Notre Dame, ⁴Department of Biological Sciences,University of Notre Dame, ⁵Department of Entomology,Kansas State University

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Overview

This article describes a method for inhibiting gene function in disease vector mosquitoes using chitosan/interfering RNA nanoparticles. The nanoparticles are ingested by mosquito larvae, allowing for gene silencing studies.

Key Study Components

Area of Science

Genetics
Entomology
Molecular Biology

Background

Gene silencing is a crucial technique in studying gene function.
Mosquitoes are significant vectors for various diseases.
Utilizing nanoparticles can enhance the delivery of RNA interference.
Chitosan is a biocompatible material suitable for nanoparticle formulation.

Purpose of Study

To develop a method for gene silencing in mosquito larvae.
To assess the effectiveness of chitosan/interfering RNA nanoparticles.
To observe the phenotypic effects of gene silencing.

Methods Used

Preparation of chitosan/interfering RNA nanoparticles.
Mixing nanoparticles with larval food to create gel pellets.
Feeding the nanoparticle-containing pellets to mosquito larvae.
Verification of gene silencing using quantitative real-time PCR and hybridization assays.

Main Results

Successful preparation of nanoparticles for gene silencing.
Observation of larvae feeding on nanoparticle pellets.
Verification of gene silencing through molecular assays.
Identification of loss of function phenotypes in treated larvae.

Conclusions

The method effectively silences gene function in mosquito larvae.
Chitosan/interfering RNA nanoparticles are a viable tool for genetic studies.
This approach can aid in understanding gene roles in disease vectors.

Frequently Asked Questions

What are chitosan/interfering RNA nanoparticles?

They are nanoparticles made from chitosan and RNA designed for gene silencing.

How are the nanoparticles administered to mosquito larvae?

They are mixed with larval food to create gel pellets that the larvae consume.

What is the purpose of gene silencing in mosquitoes?

To study the function of specific genes and their roles in disease transmission.

What methods are used to verify gene silencing?

Quantitative real-time PCR and hybridization assays are employed for verification.

What are the implications of this research?

It can lead to better understanding and control of disease vector populations.

Can this method be applied to other organisms?

While this study focuses on mosquitoes, similar techniques may be adapted for other species.

Nous décrivons ici une procédure pour inhiber la fonction des gènes chez les moustiques vecteurs de la maladie grâce à l'utilisation de chitosane / interférer nanoparticules d'ARN qui sont ingérés par les larves.

L’objectif global de cette procédure est de concevoir et d’utiliser des nanoparticules d’ARN interférentes avec les tussins pour des études de silençage génique et des larves de moustiques. Ceci est accompli en préparant d’abord des nanoparticules d’ARN interférentes avec le tussin par la combinaison de la chitine et de l’ARN interférent, suivie d’un vortex chauffant et d’une centrifugation. Le prochain objectif est de mélanger les nanoparticules avec de la nourriture et des larves pour préparer des aliments et des granulés contenant des nanoparticules contenant des granulés de gel.

La nourriture à base de nanoparticules est donnée aux larves de moustiques et les larves se nourrissant de la pastille sont observées en fin de compte après vérification du silençage génique par réaction en chaîne par polymérase quantitative en temps réel. Ou en C, deux essais d’hybridation. Les phénotypes de perte de fonction peuvent être examinés.

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