Intramolecular ligation method (iLIME) for pre-miRNA quantification and sequencing.

Minh Ngoc Le, Cong Truc Le, Tuan Anh Nguyen

RNA (New York, N.Y.) 2022 Jul

filter terms:

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. © 2022 Le et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

Citation

Minh Ngoc Le, Cong Truc Le, Tuan Anh Nguyen. Intramolecular ligation method (iLIME) for pre-miRNA quantification and sequencing. RNA (New York, N.Y.). 2022 Jul;28(7):1028-1038