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Experimental RNAi02:15

Experimental RNAi

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...
RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
RNA Interference01:23

RNA Interference

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.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...

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Related Experiment Video

Updated: Jun 2, 2026

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA
13:00

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

Published on: December 2, 2009

A comparison of multiple shRNA expression methods for combinatorial RNAi.

Glen J McIntyre1, Allison J Arndt, Kirsten M Gillespie

  • 1Johnson and Johnson Research Pty Ltd, Level 4 Biomedical Building, 1 Central Avenue, Australian Technology Park, Eveleigh, NSW, 1430, Australia. glen@madebyglen.com.

Genetic Vaccines and Therapy
|April 19, 2011
PubMed
Summary
This summary is machine-generated.

Multiple short hairpin RNAs (shRNAs) are vital for HIV-1 RNAi gene therapies. Multiple expression cassettes offer a versatile and effective strategy for co-expressing multiple shRNAs against resistant HIV-1 strains.

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Area of Science:

  • Molecular Biology
  • Gene Therapy
  • Virology

Background:

  • RNA interference (RNAi) gene therapies are promising for HIV-1 treatment.
  • Combating drug-resistant HIV-1 strains necessitates the use of multiple short hairpin RNAs (shRNAs).

Purpose of the Study:

  • To evaluate three distinct shRNA co-expression methods for their suitability in developing HIV-1 RNAi gene therapies.
  • To identify the most effective and adaptable strategy for delivering multiple shRNAs against HIV-1.

Main Methods:

  • Comparative analysis of three shRNA co-expression strategies: multiple expression vectors, multiple expression cassettes, and single transcripts with multiple dsRNA domains.
  • Assessing the efficacy and activity of different shRNA configurations against HIV-1 targets.

Main Results:

  • The multiple vector strategy showed effectiveness with two shRNAs but is limited by the number of vectors required.
  • Single transcript configurations demonstrated limited success, with only one of ten variants showing adequate activity.
  • Multiple cassette combinations (2, 3, and 4 shRNAs) were successful, maintaining activity across all positions and comparable net activities to single shRNAs.

Conclusions:

  • The multiple cassette strategy is the most suitable for current HIV-1 RNAi gene therapy development due to its ease of design, assembly, and compatibility with existing shRNA technologies.
  • This method offers a scalable and reliable approach for generating multi-shRNA therapeutics against evolving HIV-1 strains.