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Updated: Jul 18, 2026
08:53
A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
Published on: September 15, 2021
Single molecule imaging of mRNA splicing
Yo Ishihama1, Hisashi Tadakuma, Tokio Tani
1Graduate School of Science and Engineering, Waseda University, Shinjyuku-ku, Tokyo 169-8555, Japan.
Nucleic Acids Symposium Series (2004)
|December 8, 2006
Summary
Researchers developed a novel single molecule imaging assay to study pre-mRNA splicing kinetics. This technique offers new insights into the precise removal of introns, crucial for creating mature messenger RNA (mRNA).
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Eukaryotic messenger RNA precursors (pre-mRNAs) contain non-coding introns that require precise removal.
- Pre-mRNA splicing occurs within complex ribonucleoprotein machinery assembled stepwise.
- While protein factors are known to be crucial for splicing fidelity, the detailed mechanisms remain unclear.
Purpose of the Study:
- To investigate the kinetics of the pre-mRNA splicing reaction.
- To develop and utilize a novel assay system for studying splicing dynamics.
Main Methods:
- Development of a splicing assay system.
- Application of single molecule imaging techniques.
- Kinetic analysis of the splicing reaction.
Main Results:
- Established a functional splicing assay using single molecule imaging.
- Enabled real-time observation of splicing events.
- Provided kinetic data on the splicing process.
Conclusions:
- Single molecule imaging is a powerful tool for dissecting splicing mechanisms.
- The developed assay system facilitates detailed kinetic studies of pre-mRNA splicing.
- Further research using this system can elucidate the roles of protein factors in splicing fidelity.
Related Concept Videos
01:32RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
01:32RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
02:18Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
02:18Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
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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...