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pre-mRNA Processing

pre-mRNA Processing

In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...

Pre-mRNA Processing

Pre-mRNA Processing

In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps 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...

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...

Pre-mRNA Processing: Modification of pre-mRNA Ends

Pre-mRNA Processing: Modification of pre-mRNA Ends

In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps the cell...

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Related Experiment Video

Updated: Jun 19, 2026

mirMachine: A One-Stop Shop for Plant miRNA Annotation

mirMachine: A One-Stop Shop for Plant miRNA Annotation

Published on: May 1, 2021

Processing of miRNA precursors.

Yukio Kurihara¹, Yuichiro Watanabe

¹Department of Life Sciences, University of Tokyo, Tokyo, Japan.

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

|October 6, 2009

Summary

This summary is machine-generated.

This study details methods for analyzing plant microRNA (miRNA) precursor processing. These techniques help understand how Dicer-like (DCL) proteins and double-stranded RNA-binding (DRB) proteins interact with precursor RNAs.

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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

Area of Science:

Molecular Biology
Plant Science
Biochemistry

Background:

Plant microRNA (miRNA) biogenesis involves sequential cleavage of precursor molecules.
The RNase III enzyme Dicer-like protein 1 (DCL1) and HYL1 protein are known to be involved in miRNA precursor processing.
Plant miRNA processing exhibits distinct features compared to animal miRNA pathways.

Purpose of the Study:

To introduce novel methods for analyzing plant miRNA precursor processing.
To investigate the roles of DCL and DRB proteins in substrate recognition and processing.
To elucidate the mechanisms underlying differential miRNA processing in plants.

Main Methods:

Transient expression of mutated pri-miRNAs with modified stem-loop structures in Nicotiana benthamiana.
Utilizing Northern blot analysis to assess processing intermediates and products.
Employing molecular biology techniques to engineer and analyze pri-miRNA variants.

Main Results:

Demonstrated the utility of transient expression systems for studying miRNA processing.
Provided insights into how structural modifications of pri-miRNAs affect processing efficiency.
Highlighted the importance of specific structural elements in substrate recognition by DCL and DRB proteins.

Conclusions:

The developed methods offer a valuable tool for dissecting plant miRNA biogenesis.
Understanding DCL-DRB protein interactions with precursor RNAs is crucial for accurate miRNA production.
These findings contribute to a deeper comprehension of plant gene regulation via miRNAs.