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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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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...

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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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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...

RNA Interference

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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.
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Related Experiment Video

Updated: May 19, 2026

Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells

Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells

Published on: March 9, 2012

Spatiotemporal control of microRNA function using light-activated antagomirs.

Colleen M Connelly¹, Rajendra Uprety, James Hemphill

¹Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA.

Molecular Biosystems

|September 5, 2012

Summary

Researchers developed light-activated miRNA antagomirs for precise control over microRNA (miRNA) function. This photochemical method allows for unprecedented spatial and temporal resolution in studying miRNA networks and their roles in biological processes and diseases.

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Combining Optogenetics with Artificial microRNAs to Characterize the Effects of Gene Knockdown on Presynaptic Function within Intact Neuronal Circuits

Published on: March 14, 2018

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

MicroRNAs (miRNAs) are key post-transcriptional regulators involved in crucial biological processes like development and cell differentiation.
Dysregulated miRNA expression is implicated in various diseases, particularly cancer, highlighting their role in cellular networks.
Existing methods for controlling miRNA function lack the spatial and temporal precision needed for detailed functional studies.

Purpose of the Study:

To develop a novel method for precise spatiotemporal control over miRNA function using light-activated antagomirs.
To investigate the potential of photochemical regulation for studying miRNA biogenesis, biological roles, and therapeutic applications.
To enable precise manipulation of miRNA networks with high resolution in mammalian cells.

Main Methods:

Engineered miRNA antagomirs by site-specifically installing photochemically labile caging groups on selected nucleotides.
Demonstrated complete inhibition of antagomir-miRNA hybridization and function through nucleotide "caging" with restoration upon UV irradiation.
Applied caged antagomirs in mammalian cells to achieve photochemical and localized UV-induced regulation of miRNA activity.

Main Results:

Successfully synthesized light-activated antagomirs that inhibit specific microRNAs (miR-122, miR-21).
Achieved full restoration of antagomir inhibitory activity upon brief UV irradiation, demonstrating effective "decaging".
Showcased precise spatial and temporal control over miRNA function in mammalian cells via localized UV exposure.

Conclusions:

The developed photochemical approach provides unprecedented spatial and temporal resolution for regulating miRNA function and networks.
This method offers a powerful tool for investigating the roles of specific miRNAs in biological systems and disease.
The technology is adaptable for any miRNA of interest, paving the way for advanced miRNA research and potential therapeutics.