Sequencing of endogenous pre-miRNAs by iLIME-seq. (A) iLIME-seq was conducted for many endogenous pre-miRNAs in HCT116 and HEK293T cells. The sequences resulting from NGS were mapped to pre-miRNAs. Three common types of pre-miRNAs, namely intact, elongated, and trimmed, were identified. (B) iLIME-seq revealed different pre-miRNA isoforms. The 5′-end and 3′-end positions of pre-miRNAs are shown. The ac-pre-let-7, Ago2-cleaved pre-let-7, is the cleavage product of pre-let-7 by Ago2 (Diederichs and Haber 2007). (C) Diagrams to show pri-miRNAs and the positions of the 5′-end and 3′-end of their corresponding pre-miRNA isoforms. (D) The percentages of different pre-mir-142 and pre-mir-342 isoform sequences are shown as representative examples. Primer sequences used for the pre-miRNA sequencing library are shown in Supplemental Table S2.
