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Updated: Jul 20, 2026
09:29
A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
Published on: August 21, 2019
Structure analysis of microRNA precursors
Jacek Krol1, Wlodzimierz J Krzyzosiak
1Laboratory of Cancer Genetics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego, Poznan, Poland.
Methods in Molecular Biology (Clifton, N.J.)
|September 8, 2006
Summary
Investigating microRNA precursor structures reveals how variations in hairpin architecture influence microRNA liberation. Biochemical methods using nucleases and metal ions provide insights into this crucial biogenesis step.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- MicroRNA (miRNA) biogenesis involves processing precursor molecules with hairpin structures.
- The structural variability of these precursors, including loop size and stem motifs, can impact miRNA liberation efficiency.
- Understanding the structural basis of miRNA precursor processing is essential for comprehending miRNA biogenesis.
Purpose of the Study:
- To investigate the structural basis of microRNA precursor processing.
- To elucidate how precursor architecture influences microRNA liberation.
- To present biochemical methods for analyzing microRNA precursor structures.
Main Methods:
- Utilizing various nucleases and metal ions for structural analysis of microRNA precursors.
- Designing DNA template-phage promoter fusions to generate natural precursor termini.
- Performing quality control tests for sequence and structural homogeneity of in vitro transcripts before structural probing.
Main Results:
- Demonstrated protocols for analyzing microRNA precursor structures using biochemical approaches.
- Established methods for generating natural precursor ends via DNA template-phage promoter fusions.
- Validated techniques for assessing the homogeneity of in vitro transcribed precursors.
Conclusions:
- Biochemical analyses, including nuclease and metal ion probing, are valuable for studying microRNA precursor structures.
- Precise generation of precursor termini and assessment of transcript homogeneity are critical for reliable structural studies.
- These methods provide a foundation for understanding the structural determinants of microRNA liberation.
Related Concept Videos
01:22MicroRNAs
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...
01:22MicroRNAs
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...
01:22MicroRNAs
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...
02:01pre-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...
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...
02:01Pre-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...
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...
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Most of the mitochondrial precursors...
Most of the mitochondrial precursors...