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10.1261-rna.079101.122 |
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|a 13558382 (ISSN)
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|a Intramolecular ligation method (iLIME) for pre-miRNA quantification and sequencing
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|b Cold Spring Harbor Laboratory Press
|c 2022
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|a Hairpin-containing pre-miRNAs, produced from pri-miRNAs, are precursors of miRNAs (microRNAs) that play essential roles in gene expression and various human diseases. Current qPCR-based methods used to quantify pre-miRNAs are not effective to discriminate between pre-miRNAs and their parental pri-miRNAs. Here, we developed the intramolecular ligation method (iLIME) to quantify and sequence pre-miRNAs specifically. This method utilizes T4 RNA ligase 1 to convert pre-miRNAs into circularized RNAs, allowing us to design PCR primers to quantify pre-miRNAs, but not their parental primiRNAs. In addition, the iLIME also enables us to sequence the ends of pre-miRNAs using next-generation sequencing. Therefore, this method offers a simple and effective way to quantify and sequence pre-miRNAs, so it will be highly beneficial for investigating pre-miRNAs when addressing research questions and medical applications. © 2022 Le et al.
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|a high throughput sequencing
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|a High-Throughput Nucleotide Sequencing
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|a human
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|a Humans
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|a metabolism
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|a microRNA
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|a microRNA
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|a MicroRNAs
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|a pre-miRNA
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|a pre-miRNA quantification
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|a pre-miRNA sequencing
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|a pri-miRNA
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|a real time polymerase chain reaction
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|a Real-Time Polymerase Chain Reaction
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|a Le, C.T.
|e author
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|a Le, M.N.
|e author
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|a Nguyen, T.A.
|e author
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|t RNA
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|z View Fulltext in Publisher
|u https://doi.org/10.1261/rna.079101.122
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