MiR-2 family targetsawdandfngto regulate wing morphogenesis inBombyx mori

Abstract

(2015) MiR-2 family targets awd and fng to regulate wing morphogenesis in Bombyxmori, RNA Biology, 12:7, 742-748, DOI: 10.1080/15476286.2015 Keywords: CRISPR/Cas9, Gal4/UAS, miR-2 cluster, transgene, wing disc MicroRNAs (miRNAs) are post-transcriptional regulators that target specific mRNAs for repression and thus play key roles in many biological processes, including insect wing morphogenesis. miR-2 is an invertebrate-specific miRNA family that has been predicted in the fruit fly, Drosophila melanogaster, to be involved in regulating the Notch signaling pathway. We show here that miR-2 plays a critical role in wing morphogenesis in the silkworm, Bombyx mori, a lepidopteran model insect. Transgenic over-expression of a miR-2 cluster using a Gal4/UAS system results in deformed adult wings, supporting the conclusion that miR-2 regulates functions essential for normal wing morphogenesis. Two genes, abnormal wing disc (awd) and fringe (fng), which are positive regulators in Notch signaling, are identified as miR-2 targets and validated by a dual-luciferase reporter assay. The relative abundance of both awd and fng expression products was reduced significantly in transgenic animals, implicating them in the abnormal wing phenotype. Furthermore, somatic mutagenesis analysis of awd and fng using the CRISPR/Cas9 system and knock-out mutants also resulted in deformed wings similar to those observed in the miR-2 overexpression transgenic animals. The critical role of miR-2 in Bombyx wing morphogenesis may provide a potential target in future lepidopteran pest control.

Bombyx mori histone methyltransferase BmAsh2 is essential for silkworm piRNA-mediated sex determination

“…PiRNA sequences were found by referring to Kawaoka et al . [ 11 ], and the relative expression was measured using the stem-loop method [ 62 ].…”

BmHpo mutation induces smaller body size and late stage larval lethality in the silkworm, Bombyx mori

“…For example, Juvenile hormone esterase ( JHE ) was deleted in silkworms to extend the larval growth period, resulting in larger larvae and cocoons, which results in more silk production (Zhang et al ., ). In another example, knockout of abnormal wing disc ( awd ) and fringe ( fng ) resulted in deformed wings; these gene products are potential targets for control of lepidopteran pests (Ling et al ., ). Our study of the silkworm, which is a model lepidopteran insect, suggests that inhibition of the Hippo pathway should be investigated as a means for control of lepidopteran pests.…”

Comparative identification of microRNAs inApis cerana ceranaworkers’ midguts responding toNosema ceranaeinvasion

“…It has also been shown to participate in the regulation of apoptosis of neuroblasts during the normal development of the nervous system in Drosophila 70 . Recent evidence shows that miR-2 regulates functions essential for the normal wing morphogenesis of B. mori via targeting of awd and fng 71 . Alexandra’s group revealed that miRs-2/13, including miR-2a, miR-2b, miR-13a, and miR-13b, play an important regulatory part in the embryonic development of Drosophila by targeting nine genes, including Sos and Myd88 72 .…”