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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...

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...

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...

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...

Small interfering RNAs (siRNA)

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...

MicroRNAs

MicroRNAs

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 ends...

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Related Experiment Video

Updated: Jul 7, 2026

Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages

Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages

Published on: November 3, 2014

Modifications in small interfering RNA that separate immunostimulation from RNA interference.

Florian Eberle¹, Kerstin Giessler, Christopher Deck

¹Department of Medical Microbiology and Hygiene, University of Heidelberg, Heidelberg, Germany.

Journal of Immunology (Baltimore, Md. : 1950)

|February 23, 2008

Summary

Chemical modifications to synthetic small interfering RNA (siRNA) can reduce immune stimulation without compromising gene silencing. Thymidine substitutions effectively decrease type I interferon production, enhancing siRNA safety for therapeutic applications.

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MISSION esiRNA for RNAi Screening in Mammalian Cells

MISSION esiRNA for RNAi Screening in Mammalian Cells

Published on: May 12, 2010

Area of Science:

Molecular Biology
Immunology
RNA Therapeutics

Background:

Synthetic small interfering RNA (siRNA) silences gene expression via RNA interference.
siRNA can trigger type I interferon (IFN) production, leading to unwanted off-target effects.
Modifications are needed to separate immunostimulation from gene silencing activity.

Purpose of the Study:

To design chemical modifications for RNA strands to reduce immunostimulation caused by siRNA.
To investigate the impact of specific nucleobase and ribose modifications on type I IFN production and gene silencing.
To maintain siRNA potency while minimizing immune responses.

Main Methods:

Synthesized RNA strands with various chemical modifications, including 2'-deoxyuridine, thymidine, 2'-O-methyl groups, 5-methylcytidine, and 7-deazaguanosine.
Stimulated human peripheral blood mononuclear cells (PBMCs) in vitro to assess type I IFN production.
Evaluated gene-silencing activity of modified siRNA duplexes.
Utilized structure prediction and UV-melting analyses.

Main Results:

Thymidine substitutions in siRNA reduced type I IFN production without affecting gene silencing.
2'-O-methyl groups on adenosine and uridine residues decreased both IFN-alpha secretion and gene silencing.
Oligoribonucleotides with 2'-O-methyladenosine inhibited IFN-alpha secretion induced by other RNAs.
5-methylcytidine and 7-deazaguanosine residues did not affect IFN-alpha secretion.
Evidence suggests immunostimulatory sequences adopt folded structures.

Conclusions:

Chemical modifications, particularly thymidine substitutions, can suppress siRNA-induced immunostimulation.
These modifications can be introduced without compromising the gene-silencing efficacy of siRNA.
Strategic structural changes offer a method to enhance the safety and therapeutic potential of siRNA technology.