期刊
RNA
卷 28, 期 7, 页码 1028-1038出版社
COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1261/rna.079101.122
关键词
microRNA; pre-miRNA; pre-miRNA quantification; pre-miRNA sequencing; pri-miRNA
资金
- Croucher Foundation [CIA17SC03]
This article introduces a new method, intramolecular ligation method (iLIME), for quantifying and sequencing hairpin-containing pre-miRNAs. The method converts pre-miRNAs into circularized RNAs using T4 RNA ligase 1, allowing for PCR primer design for quantification and next-generation sequencing for end sequencing. This method offers a simple and effective solution for studying pre-miRNAs.
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 pri-miRNAs. 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.
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