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Interferencia de ARN larvario en el gusano de seda Bombyx mori a través de la entrega de n...
Interferencia de ARN larvario en el gusano de seda Bombyx mori a través de la entrega de n...
JoVE Journal
Biology
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JoVE Journal Biology
Larval RNA Interference in Silkworm Bombyx mori through Chitosan/dsRNA Nanoparticle Delivery

Interferencia de ARN larvario en el gusano de seda Bombyx mori a través de la entrega de nanopartículas de quitosano/dsRNA

Full Text
2,637 Views
05:36 min
October 4, 2024

DOI: 10.3791/67360-v

Jisheng Liu1, Qiangjun Yang1, Yang Yang1, Xianfeng Lin1

1School of Life Sciences,Guangzhou University

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study presents a novel protocol for the delivery of chitosan/dsRNA nanoparticles to silkworm larvae (Bombyx mori) via feeding, aimed at inducing gene silencing. The methodology is designed to facilitate gene regulation studies and high-throughput gene screening, showing significant silencing effects in targeted genes.

Key Study Components

Research Area

  • Genetic regulation
  • RNA interference (RNAi)
  • Insect model systems

Background

  • Chitosan is a biodegradable polymer that has potential in gene delivery systems.
  • Traditional RNAi methods in silkworms required injection, which is labor-intensive.
  • This feeding approach presents a cost-effective and easy alternative.

Methods Used

  • Preparation of chitosan/dsRNA nanoparticles for oral delivery.
  • Use of silkworm larvae (Bombyx mori) as a biological system.
  • Application of centrifugation and spectrophotometric methods to assess dsRNA concentration.

Main Results

  • Successful RNAi was achieved with a 79% reduction in BmToll9-2 gene transcripts.
  • Silkworm larvae treated with dsRNA were visibly smaller than control groups.
  • The cocoons from treated larvae were significantly smaller, demonstrating the effectiveness of the treatment.

Conclusions

  • The study successfully demonstrates a new method for RNAi in silkworm larvae via feeding.
  • This approach provides a labor-saving, efficient technique for genetic studies in insects.

Frequently Asked Questions

What is the significance of using chitosan/dsRNA nanoparticles?
Chitosan/dsRNA nanoparticles are biodegradable and non-toxic, making them suitable for gene delivery in insects.
How was the success of gene silencing assessed?
The level of gene silencing was quantified by measuring transcript levels of BmToll9-2.
What advantages does this feeding method offer?
It is more natural and easier than injection, reducing time and labor.
Can this method be applied to other insects?
While this study focuses on silkworms, the methodology may have potential for other insect models.
What temperatures are ideal for the nanoparticle preparation?
The solutions should be heated at 55 degrees Celsius for optimal nanoparticle formation.
How do the treated larvae differ in size?
Treated larvae are visibly smaller compared to controls, indicating effective gene silencing.
What future research could follow this study?
Future studies could investigate the effects of gene silencing on further traits or different gene targets.

Aquí se presenta un protocolo para la administración de nanopartículas de quitosano/dsRNA en larvas de gusano de seda Bombyx mori para inducir el silenciamiento génico a través de la ingestión.

El objetivo de esta investigación es desarrollar una metodología para alimentar a las larvas del gusano de seda Bombyx mori con dsRNA, resultando en un efecto de silenciamiento génico. Esto ayudará en los estudios de regulación génica y en el cribado de genes de alto rendimiento. El quitosano es un polímero barato, no tóxico y biodegradable.

Se ha descubierto que las nanopartículas de quitosano/dsRNA son eficaces para causar el efecto ARNi al alimentar a los mosquitos. En el gusano de seda, el ARNi solo se logra mediante la inyección de dsRNA hasta ahora. Sin embargo, se requieren suficientes dispositivos autenticados para la inyección.

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Interferencia de ARN larval Bombyx mori nanopartículas de quitosano/dsRNA ARN bicatenario (dsRNA) estudio funcional de genes insecto modelo de lepidópteros técnicas de ARNi alimentación de larvas de gusanos de seda sistema genético participación en la investigación en el aula

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