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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...

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...

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...

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...

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...

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Related Experiment Video

Updated: May 20, 2026

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

Published on: August 21, 2014

miRNA-like duplexes as RNAi triggers with improved specificity.

Juan G Betancur¹, Mayuko Yoda, Yukihide Tomari

¹Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo, Japan.

Frontiers in Genetics

|July 19, 2012

Summary

Small interfering RNA (siRNA) and microRNA (miRNA) pathways involve Argonaute (Ago) proteins and RNA unwinding. A new design method for artificial miRNA duplexes reduces off-target effects while maintaining silencing activity.

Keywords:

Argonaute RISC RNA-induced silencing complex asymmetry

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Last Updated: May 20, 2026

Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster

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Published on: August 21, 2014

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Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs

Published on: June 12, 2018

Area of Science:

Molecular Biology
RNA Interference
Gene Silencing

Background:

RNA interference (RNAi) relies on small RNA duplexes loaded into Argonaute (Ago) proteins.
siRNA duplexes require Ago2 slicer activity for passenger strand cleavage and unwinding.
miRNA/miRNA* duplexes utilize central mismatches for slicer-independent unwinding by any mammalian Ago protein.

Purpose of the Study:

To review current knowledge of RNA-induced silencing complex (RISC) assembly pathways.
To present a method for designing artificial miRNA/miRNA*-like duplexes.
To highlight the benefits of this design method in reducing off-target effects.

Main Methods:

Review of existing literature on RISC assembly mechanisms.
Description of a rational design approach for artificial miRNA/miRNA*-like duplexes.
Evaluation of the designed duplexes for target silencing and off-target effects.

Main Results:

Both slicer-dependent (siRNA) and slicer-independent (miRNA) unwinding mechanisms generate functional RISC.
Artificial miRNA/miRNA*-like duplexes can be designed to enhance specificity.
The proposed design method effectively reduces unwanted off-target effects.

Conclusions:

Understanding RISC assembly pathways is crucial for effective gene silencing.
Artificial miRNA design offers a strategy to improve the safety and efficacy of RNAi therapeutics.
The presented method provides a valuable tool for developing targeted gene silencing strategies.