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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.
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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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Electroporation-mediated RNA Interference Method in Odonata
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Electroporation-mediated RNA Interference Method in Odonata.

Genta Okude1, Takema Fukatsu2, Ryo Futahashi3

  • 1Department of Biological Sciences, Graduate School of Science, The University of Tokyo; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST); gentaokude@gmail.com.

Journal of Visualized Experiments : Jove
|February 22, 2021
PubMed
Summary

Researchers developed an electroporation-mediated RNA interference (RNAi) method to study gene function in dragonflies and damselflies. This technique overcomes previous limitations, enabling genetic analysis in these ancestral insects.

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Area of Science:

  • Entomology
  • Molecular Biology
  • Genetics

Background:

  • Dragonflies and damselflies (Odonata) are ancient insects with complex metamorphosis.
  • Odonata exhibit diverse coloration and well-developed vision.
  • Molecular genetic studies in Odonata are limited due to challenges with gene functional analysis, particularly RNA interference (RNAi).

Purpose of the Study:

  • To establish an effective RNA interference (RNAi) method for gene functional analysis in Odonata.
  • To overcome the inefficiency of traditional RNAi in Odonata.
  • To provide a detailed protocol for electroporation-mediated RNAi in damselflies and dragonflies.

Main Methods:

  • Developed a protocol combining in vivo electroporation with RNA interference (RNAi).
  • Detailed steps include larva preparation, dsRNA/siRNA solution preparation, injection, electroporation, and rearing.
  • Demonstrated applicability in damselflies (Ischnura senegalensis) and dragonflies (Pseudothemis zonata).

Main Results:

  • Successfully established an electroporation-mediated RNAi method for Odonata.
  • Demonstrated RNAi targeting of the melanin synthesis gene multicopper oxidase 2.
  • The method is effective in both damselflies and dragonflies.

Conclusions:

  • The electroporation-mediated RNAi protocol facilitates gene function studies in Odonata.
  • This method aids in understanding metamorphosis, morphogenesis, and color pattern formation.
  • The protocol may be applicable to other non-model organisms with low RNAi efficiency.