Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

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...
Riboswitches01:56

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
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...
siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

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.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
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...
Small interfering RNAs (siRNA)02:30

Small interfering RNAs (siRNA)

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.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Microglia-mediated protection against Alzheimer's disease pathology and detrimental effects in white matter revealed by Ptpn6 deletion.

Neuron·2026
Same author

Ionic liquids as alternative stabilizers for lipid nanoparticles used to deliver mRNA.

Philosophical transactions. Series A, Mathematical, physical, and engineering sciences·2026
Same author

Cleavage of mRNAs by a minority of pachytene piRNAs improves sperm fitness.

Nature·2026
Same author

Biochemical principles of miRNA targeting in flies.

Nature communications·2026
Same author

Chemically-inducible CRISPR/Cas9 circuits for ultra-high dynamic range gene perturbation.

Nature communications·2025
Same author

An unbiased whole-genome open reading frame overexpression screen identifies B3GALT2, a novel inducer of cellular ASO activity.

Molecular therapy. Nucleic acids·2025

Related Experiment Video

Updated: Jul 17, 2026

MISSION esiRNA for RNAi Screening in Mammalian Cells
15:31

MISSION esiRNA for RNAi Screening in Mammalian Cells

Published on: May 12, 2010

A protein sensor for siRNA asymmetry.

Yukihide Tomari1, Christian Matranga, Benjamin Haley

  • 1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA.

Science (New York, N.Y.)
|November 20, 2004
PubMed
Summary

Small interfering RNAs (siRNAs) guide RNA interference (RNAi) by unwinding and assembling into the RNA-induced silencing complex (RISC). The R2D2 protein ensures correct siRNA strand loading into RISC by sensing thermodynamic asymmetry.

More Related Videos

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster
09:39

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Published on: August 21, 2014

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids
09:04

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids

Published on: September 21, 2017

Related Experiment Videos

Last Updated: Jul 17, 2026

MISSION esiRNA for RNAi Screening in Mammalian Cells
15:31

MISSION esiRNA for RNAi Screening in Mammalian Cells

Published on: May 12, 2010

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster
09:39

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Published on: August 21, 2014

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids
09:04

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids

Published on: September 21, 2017

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • RNA interference (RNAi) is a biological process in which small interfering RNAs (siRNAs) regulate gene expression.
  • siRNAs must be processed and loaded into the RNA-induced silencing complex (RISC) to mediate gene silencing.
  • The selection of the guide strand for RISC loading is crucial for RNAi pathway specificity.

Purpose of the Study:

  • To investigate the mechanism by which siRNA strands are selected for RISC loading in Drosophila.
  • To identify the role of the Dicer-2/R2D2 protein complex in orienting siRNAs for RISC assembly.
  • To understand how thermodynamic asymmetry of siRNA duplexes influences guide strand selection.

Main Methods:

  • Biochemical assays to study protein-siRNA interactions.
  • Analysis of siRNA duplexes and their thermodynamic properties.
  • In vivo studies in Drosophila to observe RISC loading and gene silencing.

Main Results:

  • The Dicer-2/R2D2 heterodimer orients on the siRNA duplex based on binding to the end with greater double-stranded character.
  • R2D2 preferentially binds the siRNA end with a 5'-phosphate, which is excluded from the RISC.
  • This binding preference dictates which siRNA strand is incorporated into the RISC.

Conclusions:

  • R2D2 acts as a sensor for thermodynamic asymmetry in siRNA duplexes.
  • R2D2 functions as a licensing factor, ensuring that only authentic siRNAs enter the RNAi pathway.
  • The study elucidates a key regulatory step in siRNA-mediated gene silencing.