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Updated: Oct 14, 2025

mirMachine: A One-Stop Shop for Plant miRNA Annotation
Published on: May 1, 2021
miRPreM and tiRPreM: Improved methodologies for the prediction of miRNAs and tRNA-induced small non-coding RNAs for
Hukam Chand Rawal1,2, Shakir Ali2,3, Tapan Kumar Mondal1
1ICAR-National Institute for Plant Biotechnology, LBS Centre, Pusa, New Delhi 110012, India.
Abstract:
In recent years, microRNAs (miRNAs) and tRNA-derived RNA fragments (tRFs) have been reported extensively following different approaches of identification and analysis. Comprehensively analyzing the present approaches to overcome the existing variations, we developed a benchmarking methodology each for the identification of miRNAs and tRFs, termed as miRNA Prediction Methodology (miRPreM) and tRNA-induced small non-coding RNA Prediction Methodology (tiRPreM), respectively. We emphasized the use of respective genome of organism under study for mapping reads, sample data with at least two biological replicates, normalized read count support and novel miRNA prediction by two standard tools with multiple runs. The performance of these methodologies was evaluated by using Oryza coarctata, a wild rice species as a case study for model and non-model organisms. With organism-specific reference genome approach, 98 miRNAs and 60 tRFs were exclusively found. We observed high accuracy (13 out of 15) when tested these genome-specific miRNAs in support of analyzing the data with respective organism. Such a strong impact of miRPreM, we have predicted more than double number of miRNAs (186) as compared with the traditional approaches (79) and with tiRPreM, we have predicted all known classes of tRFs within the same small RNA data. Moreover, the methodologies presented here are in standard form in order to extend its applicability to different organisms rather than restricting to plants. Hence, miRPreM and tiRPreM can fulfill the need of a comprehensive methodology for miRNA prediction and tRF identification, respectively, for model and non-model organisms.
Insights
We developed new methods, miRPreM and tiRPreM, for accurately identifying microRNAs (miRNAs) and tRNA-derived RNA fragments (tRFs). These benchmarking tools improve small RNA analysis in both model and non-model organisms.
Area of Science:
- Molecular Biology
- Genomics
- Bioinformatics
Background:
- MicroRNAs (miRNAs) and tRNA-derived RNA fragments (tRFs) are crucial small non-coding RNAs with diverse biological roles.
- Existing methods for identifying miRNAs and tRFs often yield variable results due to differing analytical approaches.
Purpose of the Study:
- To develop and validate robust benchmarking methodologies for the accurate identification of miRNAs (miRPreM) and tRFs (tiRPreM).
- To enhance the reliability and comparability of small non-coding RNA analysis across different organisms.
Main Methods:
- Developed miRPreM and tiRPreM, emphasizing organism-specific genome mapping, biological replicates, normalized read counts, and dual tool validation for novel miRNA prediction.
- Applied the methodologies to Oryza coarctata (wild rice) as a case study for both model and non-model organisms.
Main Results:
- The organism-specific approach identified 98 unique miRNAs and 60 tRFs in Oryza coarctata.
- miRPreM identified over double the number of miRNAs (186) compared to traditional methods (79), with high accuracy.
- tiRPreM successfully identified all known classes of tRFs within the analyzed small RNA data.
Conclusions:
- miRPreM and tiRPreM provide a standardized and comprehensive approach for miRNA and tRF identification.
- These methodologies are applicable to a wide range of organisms, including plants, animals, and potentially others.
- The developed tools address existing variations in small RNA analysis, improving accuracy and discovery potential.
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