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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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siRNA - Small Interfering RNAs02:30

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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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MicroRNAs01:22

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae
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Gene suppression in schistosomes using RNAi.

Akram A Da'dara1, Patrick J Skelly

  • 1Molecular Helminthology Laboratory, Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 13, 2014
PubMed
Summary

Schistosomiasis, a deadly tropical disease, faces drug resistance challenges. RNA interference (RNAi) offers a new method to explore gene targets for novel treatments against Schistosoma mansoni parasites.

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Area of Science:

  • Parasitology
  • Molecular Biology
  • Genetics

Background:

  • Schistosomiasis is a major global health concern caused by parasitic worms, leading to over 300,000 deaths annually.
  • Current praziquantel treatment for schistosomiasis is threatened by emerging drug resistance.
  • Advancements in genomic sequencing of schistosome species offer opportunities for developing new interventions.

Purpose of the Study:

  • To review methods for applying RNA interference (RNAi) to suppress gene expression in Schistosoma mansoni.
  • To validate RNAi as a tool for exploring drug targets and understanding parasite biology.

Main Methods:

  • Detailed procedures for isolating and culturing intra-mammalian life stages (adults and schistosomula) of Schistosoma mansoni.
  • Methods for preparing and delivering double-stranded RNA (dsRNA) to target specific genes.
  • Quantitative real-time PCR (qPCR) for evaluating gene suppression efficacy.

Main Results:

  • RNA interference (RNAi) is effective in suppressing gene expression across tested parasite life stages.
  • Established protocols enable gene-specific knockdown in adult and schistosomula stages of Schistosoma mansoni.
  • Quantitative real-time PCR provides a reliable method for assessing gene suppression.

Conclusions:

  • RNAi is a valuable tool for functional genomics and drug target validation in Schistosoma mansoni.
  • The described methods facilitate research into novel schistosomiasis treatments.
  • Further application of RNAi can advance our understanding of schistosome biology and control strategies.