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

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...
Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
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...
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...
Ribozymes02:47

Ribozymes

The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
piRNA - Piwi-interacting RNAs02:57

piRNA - Piwi-interacting RNAs

PIWI-interacting RNAs, or piRNAs, are the most abundant short non-coding RNAs. More than 20,000 genes have been found in humans that code for piRNAs while only 2000 genes have been found for miRNAs. piRNAs can act at the transcriptional and post-transcriptional levels and have a vital role in silencing transposable elements present in germ cells. They are also involved in epigenetic silencing and activation. Previously, they were thought to function only in germ cells but new evidence suggests...

You might also read

Related Articles

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

Sort by
Same author

Programming T cells for intercellular genome editing.

bioRxiv : the preprint server for biology·2026
Same author

Harmonizing standards and resources for the medical genome.

Nature·2026
Same author

Publisher Correction: Lung and liver editing by lipid nanoparticle delivery of a stable CRISPR-Cas9 ribonucleoprotein.

Nature biotechnology·2026
Same author

Targeting cancer-specific mutations with RNA-triggered chromatin shredding.

Nature·2026
Same author

Structural basis of RNA-guided DNA integration by type I CRISPR-associated transposases.

bioRxiv : the preprint server for biology·2026
Same author

Temperate phages enhance bacterial host fitness via RNA-guided flagellar remodelling.

Nature microbiology·2026

Related Experiment Video

Updated: Jun 8, 2026

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
09:16

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro

Published on: May 3, 2014

Substrate-specific kinetics of Dicer-catalyzed RNA processing.

Srinivas Chakravarthy1, Samuel H Sternberg, Colleen A Kellenberger

  • 1Howard Hughes Medical Institute, Berkeley, CA 94720, USA.

Journal of Molecular Biology
|October 12, 2010
PubMed
Summary

Human Dicer enzyme processes microRNA precursors much faster than short interfering RNA precursors. The trans-activation response RNA binding protein (TRBP) enhances this RNA processing for both types of substrates.

More Related Videos

Extremely Rapid and Specific Metabolic Labelling of RNA In Vivo with 4-Thiouracil (Ers4tU)
11:46

Extremely Rapid and Specific Metabolic Labelling of RNA In Vivo with 4-Thiouracil (Ers4tU)

Published on: August 22, 2019

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle
10:05

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle

Published on: March 5, 2019

Related Experiment Videos

Last Updated: Jun 8, 2026

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
09:16

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro

Published on: May 3, 2014

Extremely Rapid and Specific Metabolic Labelling of RNA In Vivo with 4-Thiouracil (Ers4tU)
11:46

Extremely Rapid and Specific Metabolic Labelling of RNA In Vivo with 4-Thiouracil (Ers4tU)

Published on: August 22, 2019

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle
10:05

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle

Published on: March 5, 2019

Area of Science:

  • Molecular Biology
  • RNA Biology
  • Gene Regulation

Background:

  • Dicer is a key enzyme in eukaryotic gene expression, producing microRNAs (miRNAs) and short interfering RNAs (siRNAs).
  • Dicer processes double-stranded RNA (dsRNA) into small regulatory RNAs.
  • The role of dsRNA binding proteins, like trans-activation response (TAR) RNA binding protein (TRBP), in Dicer substrate selection and efficiency is not fully understood.

Purpose of the Study:

  • To investigate the mechanistic differences between Dicer processing of pre-miRNA and pre-siRNA substrates.
  • To determine how TRBP influences substrate selection and RNA processing efficiency by Dicer.

Main Methods:

  • Enzymatic assays using pre-miRNA and pre-siRNA substrates.
  • Michaelis-Menten kinetic analysis to determine maximal cleavage rates (Vmax).
  • Investigated the role of TRBP's dsRNA binding domains in Dicer activity.

Main Results:

  • Human Dicer exhibits significantly faster processing of pre-miRNA compared to pre-siRNA substrates.
  • Maximal cleavage rates (Vmax) for pre-miRNA were over 100-fold higher than for pre-siRNA.
  • TRBP enhanced the dicing of both substrates, requiring its N-terminal dsRNA binding domains.

Conclusions:

  • Dicer substrate processing kinetics are influenced by substrate structure.
  • TRBP enhances Dicer activity by stabilizing Dicer-substrate complexes.
  • Dicer's product RNA generation rate is partly determined by substrate structural properties.