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

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...

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...

You might also read

Related Articles

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

Sort by

Same author

SIRT1 rs10823108 and FOXO1 rs17446614 responsible for genetic susceptibility to diabetic nephropathy.

Scientific reports·2017

Same author

Repetitive Transcranial Magnetic Stimulation Ameliorates Cognitive Impairment by Enhancing Neurogenesis and Suppressing Apoptosis in the Hippocampus in Rats with Ischemic Stroke.

Frontiers in physiology·2017

Same author

GSK-3Beta-Dependent Activation of GEF-H1/ROCK Signaling Promotes LPS-Induced Lung Vascular Endothelial Barrier Dysfunction and Acute Lung Injury.

Frontiers in cellular and infection microbiology·2017

Same author

Decreased high density lipoprotein cholesterol is an independent predictor for persistent organ failure, pancreatic necrosis and mortality in acute pancreatitis.

Scientific reports·2017

Same author

Pancreatic neuroendocrine neoplasms: Correlation between MR features and pathological tumor grades.

Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban·2017

Same author

Two new monoterpenoid indole alkaloids from Alstonia rostrata.

Natural product research·2017

Search research articles

Related Experiment Video

Updated: May 6, 2026

A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

Published on: August 21, 2019

Primary microRNA processing assay reconstituted using recombinant Drosha and DGCR8.

Ian Barr¹, Feng Guo

¹Department of Biological Chemistry, University of California Los Angeles, Los Angeles, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)

|October 30, 2013

Summary

Researchers reconstituted microRNA (miRNA) processing using recombinant human Drosha and DiGeorge critical region 8 (DGCR8) proteins. DGCR8, bound to an Fe(III) heme cofactor, is crucial for pri-miRNA cleavage activity.

More Related Videos

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Published on: August 3, 2011

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

Published on: June 16, 2022

Related Experiment Videos

Last Updated: May 6, 2026

A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

Published on: August 21, 2019

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Published on: August 3, 2011

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

Published on: June 16, 2022

Area of Science:

Biochemistry
Molecular Biology
Gene Regulation

Background:

The Microprocessor complex, comprising Drosha and DGCR8, processes primary microRNAs (pri-miRNAs) into precursor miRNAs in the nucleus.
DGCR8 binds an Fe(III) heme cofactor, which is essential for activating its catalytic activity in pri-miRNA processing.

Purpose of the Study:

To describe the reconstitution of pri-miRNA processing using recombinant human Drosha and DGCR8 proteins.
To detail methods for expressing and purifying Fe(III) heme-bound DGCR8, the most active form.

Main Methods:

Expression and purification of recombinant human Drosha and DGCR8 proteins.
Reconstitution of the Microprocessor complex for in vitro pri-miRNA processing assays.
Quantification of Fe(III) heme content in purified DGCR8.

Main Results:

Successful reconstitution of pri-miRNA processing activity using recombinant proteins.
Established procedures for obtaining active, heme-bound DGCR8.
Characterization of the Fe(III) heme-DGCR8 dimer as the most catalytically potent form.

Conclusions:

Recombinant human Drosha and DGCR8 can effectively reconstitute pri-miRNA processing.
The Fe(III) heme-bound DGCR8 dimer is key to Microprocessor complex activity.
This work provides a foundation for further mechanistic studies of miRNA biogenesis.