Over-expression of RNA interference (RNAi) core machinery improves susceptibility to RNAi in silkworm larvae

L. You, F. Zhang, S. Huang, A. Merchant, X. Zhou, Z. Li

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

RNA interference (RNAi), one of the strategies that organisms use to defend against invading viruses, is an important tool for functional genomic analysis. In insects, the efficacy of RNAi varies amongst taxa. Lepidopteran insects are, in large part, recalcitrant to RNAi. The overall goal of this study is to overcome such insensitivity in lepidopterans to RNAi. We hypothesize that over-expression of core RNAi machinery enzymes can improve RNAi efficacy in traditionally recalcitrant species. A transgenic Bombyx mori strain, Baculovirus Immediate-Early Gene, ie1, promoter driven expression of silkworm Dicer2 coding sequence (IE1-BmDicer2), which over-expresses BmDicer2, was generated by piggyBac transposon-mediated transgenesis. Two indexes, the ratio of animals that showed a silencing phenotype and the duration of silencing, were used to evaluate silencing efficiency. Significant knockdown of target gene expression was observed at 48 h postinjection at both the transcriptional and translational levels. Furthermore, we coexpressed B. mori Argonaute 2 BmAgo2)and BmDicer 2 and found that 22% of the animals (n = 18) showed an obvious silencing effect even at 72 h, suggesting that coexpression of these two RNAi core machinery enzymes further increased the susceptibility of B. mori to injected double-stranded RNAs. This study offers a new strategy for functional genomics research in RNAi-refractory insect taxa in general and for lepidopterans in particular.

Original languageEnglish
Pages (from-to)353-362
Number of pages10
JournalInsect Molecular Biology
Volume29
Issue number4
DOIs
StatePublished - Aug 1 2020

Bibliographical note

Publisher Copyright:
© 2020 The Royal Entomological Society

Keywords

  • Bombyx mori
  • Dicer2
  • RNAi core machinery
  • RNAi efficacy
  • transgenic silkworm

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Insect Science

Fingerprint

Dive into the research topics of 'Over-expression of RNA interference (RNAi) core machinery improves susceptibility to RNAi in silkworm larvae'. Together they form a unique fingerprint.

Cite this