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. 2018 Sep;28(9):1309–1318. doi: 10.1101/gr.233809.117

Figure 1.

Figure 1.

Emergence of de novo miRNAs and the creation of knockout (KO) lines. (A) The emergence of new miRNAs in Drosophila. The numbers in red indicate miRNA emergence in each time interval of Drosophila phylogeny. Note the “pyramid” age structure with many very young miRNAs, suggesting many deaths before aging. The age distribution is sourced from our previous study (Lyu et al. 2014). (D. mel) D. melanogaster; (D. sim) D. simulans; (D. pse) D. pseudoobscura; (D. vir) D. virilis. (B) miRNA mutants in this study fall into two de novo miRNA clusters. De novo miRNAs tend to be in clusters and are X-linked (Lyu et al. 2014). (*) The six knockout creations of the two clusters of focus. Colors denote miRNA age as indicated. (C–H) Verification of miRNA knockout efficiency. Upper: DNA sequences of miRNA KO strains with the mature sequences highlighted in yellow. Lower: relative expression of miRNAs in the testis. No miRNAs are detectable in the KO lines, whereas the expressions of the neighboring miRNAs are normal (positive controls). Note that mir-978 has two 5′ iso-miRNAs (miR-978-3p1 and miR-978-3p2), which are processed from one precursor in the D. melanogaster testis.