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Updated: Mar 9, 2026
RNA Blot Analysis for the Detection and Quantification of Plant MicroRNAs
Published on: July 11, 2020
A three-way junction structure-based isothermal exponential amplification strategy for sensitive detection of
Xiangdong Wang1, Hui Wang1, Chenghui Liu1
1Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an 710062, Shaanxi Province, P. R. China. liuch@snnu.edu.cn lzpbd@snnu.edu.cn.
Abstract:
Plant microRNAs (miRNAs) differ from animal miRNAs in that they are 2'-O-methylated at the 3' end nucleotide which limits the ability of plant miRNAs to serve as primers for polymerization reactions. Herein, we have rationally designed a three-way junction structure-based isothermal exponential amplification reaction (3WJ-EXPAR) which is well-suited for sensitive detection of 3'-terminal 2'-O-methylated miRNAs.
Insights
Plant microRNAs (miRNAs) have a unique 2'-O-methylation preventing their use in polymerization. We developed a novel three-way junction-based isothermal exponential amplification reaction (3WJ-EXPAR) for sensitive detection of these methylated miRNAs.
Area of Science:
- Biochemistry
- Molecular Biology
- Plant Science
Background:
- Plant microRNAs (miRNAs) possess a unique 3"-terminal 2"-O-methylation.
- This methylation inhibits their function as primers in polymerization reactions, posing challenges for detection.
- Existing detection methods may not be optimized for these specific modifications.
Purpose of the Study:
- To design and validate a novel amplification reaction for sensitive detection of 3"-terminal 2"-O-methylated plant miRNAs.
- To overcome the limitations imposed by the 2"-O-methylation on miRNA detection.
- To provide a new tool for studying plant miRNA biology.
Main Methods:
- Rational design of a three-way junction structure.
- Development of an isothermal exponential amplification reaction (EXPAR) system.
- Integration of the three-way junction structure into the EXPAR for specific miRNA recognition and amplification.
Main Results:
- Successful design and implementation of the three-way junction structure-based isothermal exponential amplification reaction (3WJ-EXPAR).
- Demonstrated high sensitivity in detecting 3"-terminal 2"-O-methylated plant miRNAs.
- The 3WJ-EXPAR system effectively overcomes the inhibitory effect of 2"-O-methylation.
Conclusions:
- The 3WJ-EXPAR is a powerful and sensitive method for detecting 3"-terminal 2"-O-methylated plant miRNAs.
- This method offers a significant advancement in the field of plant small RNA detection.
- The developed system has potential applications in plant research and diagnostics.
