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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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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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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 the pre-miRNA...
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Related Experiment Video

Updated: Mar 26, 2026

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae
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Gene Silencing in Insect Cells Using RNAi.

Hsuan-Chen Wu1, John C March2, William E Bentley3,4

  • 1Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan.

Methods in Molecular Biology (Clifton, N.J.)
|January 29, 2016
PubMed
Summary

This study presents a fast method for creating and delivering double-stranded RNA (dsRNA) for RNA interference (RNAi) in insect cells. The technique ensures efficient gene silencing with quick cell recovery, offering practical design guidelines.

Keywords:
Gene silencingRNA interferenceRNAiSP6T7TranscriptiondsRNAmRNA

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • RNA interference (RNAi) is a crucial gene silencing mechanism.
  • Efficient delivery of double-stranded RNA (dsRNA) is vital for RNAi applications.
  • Sf-21 cell cultures are widely used in insect cell-based research.

Purpose of the Study:

  • To describe a streamlined method for synthesizing and transfecting dsRNA in Sf-21 cells.
  • To provide guidelines and resources for effective dsRNA design.
  • To optimize the RNAi process for rapid gene silencing and cell recovery.

Main Methods:

  • Synthesis of dsRNA using essential kits (RNAqueous®-4PCR, Megascript™ T7, Superscript™ III).
  • Rapid transfection protocol for Sf-21 cell cultures, completed within one hour.
  • Inclusion of detailed methods for dsRNA design and online resource recommendations.

Main Results:

  • Successful synthesis and transfection of dsRNA in Sf-21 cells.
  • Demonstration of a one-hour transfection window.
  • Observed rapid cell recovery within 12 hours post-transfection.

Conclusions:

  • The described technique offers an efficient and rapid approach for RNA interference in Sf-21 cells.
  • The provided dsRNA design suggestions and resources facilitate effective gene silencing.
  • This method optimizes dsRNA delivery and cell viability for research applications.