Related Experiment Videos
Analysis of 3' End Modifications in microRNAs by High-Throughput Sequencing
Madalena M Reimão-Pinto1, Angela M Rodrigues-Viana1, Stefan L Ameres2
1IMBA - Institute of Molecular Biotechnology, Vienna Biocenter, Vienna, Austria.
Methods in Molecular Biology (Clifton, N.J.)
|July 1, 2018
Summary
Researchers developed new methods to systematically study microRNA tailing, a process adding nucleotides to precursor microRNAs. This research aids in understanding gene regulation and its role in development and disease.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- MicroRNAs (miRNAs) are small, non-coding RNAs regulating gene expression post-transcriptionally.
- miRNA biogenesis is controlled by mechanisms including the addition of non-templated nucleotides (tailing) to precursor miRNAs.
- Tailing, predominantly uridine addition, impacts miRNA function and regulation.
Purpose of the Study:
- To describe systematic methods for characterizing tailing events in mature and precursor microRNAs.
- To provide protocols for cDNA library preparation for analyzing untemplated nucleotide additions.
- To facilitate the study of posttranscriptional modifications in gene regulatory small RNAs.
Main Methods:
- Development of untargeted and targeted cDNA library preparation protocols.
- Application of methods to precursor-microRNAs in Drosophila melanogaster.
- Utilization of a computational framework for analyzing untemplated nucleotide additions in cDNA libraries.
Main Results:
- Successful implementation of protocols for characterizing miRNA tailing events.
- Demonstration of methods in a model organism (Drosophila melanogaster).
- Establishment of a computational framework for analyzing tailing data.
Conclusions:
- The described methods enable systematic characterization of posttranscriptional modifications in small RNAs.
- These protocols will advance the understanding of regulatory mechanisms controlling gene silencing.
- Clarification of tailing's role in miRNA biogenesis and function is anticipated.