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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...

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...

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...

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Related Experiment Video

Updated: May 16, 2026

Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library

Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library

Published on: April 6, 2012

An arrayed RNA interference genome-wide screen identifies candidate genes involved in the MicroRNA 21 biogenesis

David Shum¹, Bhavneet Bhinder, Christina N Ramirez

¹High-Throughput Screening Core Facility, Molecular Pharmacology and Chemistry Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.

Assay and Drug Development Technologies

|November 17, 2012

Summary

Researchers screened 64,755 small interfering RNAs (siRNAs) to identify genes regulating microRNA-21 (miR-21) biogenesis. This screen identified 1,273 candidate genes, offering new targets for therapeutic development.

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Last Updated: May 16, 2026

Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library

Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library

Published on: April 6, 2012

Detection of miRNA Targets in High-throughput Using the 3'LIFE Assay

Detection of miRNA Targets in High-throughput Using the 3'LIFE Assay

Published on: May 25, 2015

CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

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Published on: April 25, 2022

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

MicroRNAs (miRNAs) are crucial regulators of gene expression involved in development and differentiation.
Dysregulation of miRNAs is linked to diseases like cancer, highlighting the need to understand their biogenesis pathways.
Identifying genes and pathways controlling miRNA biogenesis is essential for therapeutic interventions.

Purpose of the Study:

To develop and validate a high-content assay for screening modulators of microRNA biogenesis.
To conduct a genome-wide screen to identify novel regulators of microRNA-21 (miR-21) biogenesis.
To nominate potential druggable targets for regulating miRNA function.

Main Methods:

Implemented a cell-based reporter assay using enhanced green fluorescent protein (EGFP) controlled by miR-21.
Integrated cell metabolic activity (Alamar Blue) and cell death (Hoechst staining) measurements.
Performed an arrayed genome-wide short interfering RNA (siRNA) screen (64,755 siRNAs) and applied a stringent analysis method (Bhinder-Djaballah).

Main Results:

Validated the assay strategy using known genes.
Successfully completed a genome-wide siRNA screen identifying 1,273 candidate genes regulating miR-21 biogenesis.
Biological classification revealed key control points, including clathrin-mediated endocytosis in vesicular transport.

Conclusions:

The study successfully identified novel candidate regulators of miR-21 biogenesis through a genome-wide screen.
The identified genes represent potential druggable targets for developing small molecules to modulate miRNA function.
This work provides a foundation for further research into miRNA regulation and therapeutic development.