What is it about?
There is often the need to determine the simultaneous presence of a miRNA and its target mRNA in a specific tissue to understand their molecular mechanisms. A method for simultaneously studying the spatial-temporal expression patterns of miRNAs and mRNAs in tissues to achieve the resolution at the single-cell level is developed.
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Why is it important?
A method that uses sequence-specific miRNA-locked nucleic acid (LNA) and mRNA-LNA probes is developed. Moreover, it augments the detection signal by rolling circle amplification, achieving a high signal-noise ratio at the single-cell level. Dot signals are counted for determining the expression levels of mRNA and miRNA molecules in specific cells. We show a high sequence specificity of our miRNA-LNA probe, revealing that it can discriminate single-base mismatches. Numerical quantification by our method is tested in transgenic rice lines with different gene expression levels.
Perspectives
We have developed a method that is capable of simultaneously detecting miRNA and mRNA molecules in plant tissues by combing LNA probes and padlock probes with RCA to achieve high sequence specificity and sequence-based signal amplification. The method is robust, versatile and scalable to meet the emerging demands for detection of gene expression at the single-cell level.
Chi-Chih Wu
Academia Sinica
Read the Original
This page is a summary of: Simultaneous detection of
miRNA
and
mRNA
at the single‐cell level in plant tissues, Plant Biotechnology Journal, September 2022, Wiley,
DOI: 10.1111/pbi.13931.
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