Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

siRNA - Small Interfering RNAs

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 RNAi

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 Interference

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...

You might also read

Related Articles

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

Sort by

Same author

KMT2A and KMT2B episignatures address diagnostic challenges associated with rare neurodevelopmental disorders.

Genetics in medicine : official journal of the American College of Medical Genetics·2026

Same author

A Homozygous Deep Intronic SNX14 Variant Activates Pseudo-Exon Inclusion in a Patient with SCAR20.

Genes·2026

Same author

Bi-allelic ATG12 variants impair autophagy and cause a neurodevelopmental disorder.

American journal of human genetics·2026

Same author

Biallelic SUPT4H1 variants cause a multisystem neurodevelopmental disorder associated with disrupted transcription.

Genetics in medicine : official journal of the American College of Medical Genetics·2026

Same author

De novo CSDE1 missense variant T595N regulates miRNA expression in a patient with a complex neurodevelopmental condition.

Biochimica et biophysica acta. Molecular basis of disease·2025

Same author

CRISPR Activation Reveals the Spliceogenicity of an Intronic NEB Variant in Fetuses With Arthrogryposis Multiplex Congenita 6.

Clinical genetics·2025

Search research articles

Related Experiment Video

Updated: Jun 16, 2026

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

shRNA expression constructs designed directly from siRNA oligonucleotide sequences.

Tuva Barøy¹, Kirsten Sørensen, Mona Mari Lindeberg

¹Institute of Medical Genetics, Faculty of Medicine, University of Oslo, PO Box 1036, Blindern, 0315 Oslo, Norway. tuva.baroy@medisin.uio.no

Molecular Biotechnology

|February 2, 2010

Summary

This study demonstrates that small interfering RNA (siRNA) sequences designed using existing algorithms can effectively silence gene expression when converted into short hairpin RNA (shRNA) constructs for stable knockdown in mammalian cells.

More Related Videos

DNA Vector-based RNA Interference to Study Gene Function in Cancer

DNA Vector-based RNA Interference to Study Gene Function in Cancer

Published on: June 4, 2012

Using In Vitro and In-cell SHAPE to Investigate Small Molecule Induced Pre-mRNA Structural Changes

Using In Vitro and In-cell SHAPE to Investigate Small Molecule Induced Pre-mRNA Structural Changes

Published on: January 30, 2019

Related Experiment Videos

Last Updated: Jun 16, 2026

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

DNA Vector-based RNA Interference to Study Gene Function in Cancer

DNA Vector-based RNA Interference to Study Gene Function in Cancer

Published on: June 4, 2012

Using In Vitro and In-cell SHAPE to Investigate Small Molecule Induced Pre-mRNA Structural Changes

Using In Vitro and In-cell SHAPE to Investigate Small Molecule Induced Pre-mRNA Structural Changes

Published on: January 30, 2019

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

RNA interference (RNAi) is a key mechanism for gene silencing.
Transient gene knockdown is achieved using small interfering RNA (siRNA).
Stable gene knockdown can be achieved using short hairpin RNA (shRNA).

Purpose of the Study:

To evaluate the efficacy of siRNA-designed sequences when used in shRNA constructs.
To determine if established siRNA design algorithms can predict efficient shRNA sequences.
To assess the gene silencing potential of shRNA constructs derived from siRNA sequences.

Main Methods:

Construction of 25 shRNA cassettes using sequences from 25 siRNAs targeting five independent transcripts.
Transfection of both siRNAs and shRNA constructs into HEK293T cells.
Quantitative real-time PCR (qRT-PCR) to measure gene expression levels.

Main Results:

19 out of 25 shRNA constructs successfully reduced average gene expression to below 30%.
The shRNA constructs derived from siRNA sequences demonstrated significant gene knockdown.
The study validated the effectiveness of converting siRNA sequences for shRNA-based gene silencing.

Conclusions:

Sequences designed by siRNA algorithms are effective for generating efficient shRNA expression constructs.
The conversion of siRNA sequences into shRNA cassettes is a viable strategy for stable gene knockdown.
This approach offers a reliable method for target-specific gene silencing in mammalian systems.