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• W1997386341 abstract "<ns4:p>microRNA (miRNA) regulation is crucial to achieve precise spatio-temporal expression patterns of their target genes. This makes it crucial to determine the levels of cleavage of a particular target mRNA in different tissues and under different conditions. We developed a quantitative PCR method “quantitative Amplification of Cleaved Ends (qACE)” to assay levels of specific cleavage products in order to determine the extent of miRNA-directed target cleavage of a specific target gene. qACE uses cDNA generated from adapter-ligated RNA molecules and relies on a carefully designed fusion primer that spans the adapter-cleaved RNA junction in qPCR to specifically amplify and quantify cleaved products. The levels of full-length transcripts can also be assayed in the same cDNA preparation using primers that span across the miRNA cleavage site. We used qACE to demonstrate that soybean roots over-expressing miR164 had increased levels of target cleavage and that miRNA deficient <ns4:italic>Arabidopsis thaliana</ns4:italic> <ns4:italic>hen1-1</ns4:italic> mutants had reduced levels of target cleavage. We used qACE to discover that differential cleavage by miR164 in nodule <ns4:italic>vs.</ns4:italic> adjacent root tissue contributed to nodule-specific expression of NAC1 transcription factors in soybean. These experiments show that qACE can be used to discover and demonstrate tissue-specific cleavage by miRNAs to achieve specific spatio-temporal expression of target genes in plants.</ns4:p>" @default.
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• W1997386341 date "2015-07-01" @default.
• W1997386341 modified "2023-09-27" @default.
• W1997386341 title "Quantitative Amplification of Cleaved Ends (qACE) to assay miRNA-directed target cleavage" @default.
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• W1997386341 doi "https://doi.org/10.12688/f1000research.5266.2" @default.
• W1997386341 hasPublicationYear "2015" @default.
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