A Random-Screening Approach to Identify RNAi Targets for the Control of Western Corn Rootworm (Diabrotica. virgifera virgifera Le Conte)

Methods Mol Biol. 2022:2360:91-103. doi: 10.1007/978-1-0716-1633-8_9.

Abstract

Identification of active target genes in bioassay screening is the first important step for application of RNA interference (RNAi) for pest control. Here, we describe the methodology for performing high-throughput RNAi target screening against important agriculture pest, Western corn rootworm in 96-well microplate. Two approaches are presented to identify active targets from random-cDNA library or testing a certain group of specific targets via in silico sequence analysis. Methods of PCR primer design, DNA template preparation, and dsRNA production described here can be applied for other pests.

Keywords: Diabrotica virgifera virgifera; Diet assay; Double-stranded RNA (dsRNA); In vitro transcription (IVT ); RNA interference (RNAi); Western corn rootworm (WCR ).

MeSH terms

  • Animals
  • Coleoptera* / genetics
  • Larva
  • RNA Interference
  • RNA, Double-Stranded
  • Zea mays* / genetics

Substances

  • RNA, Double-Stranded