Expanding the genetic toolkit of Tribolium castaneum

PLoS One. 2018 Apr 12;13(4):e0195977. doi: 10.1371/journal.pone.0195977. eCollection 2018.

Abstract

The red flour beetle, Tribolium castaneum, is an important model insect and agricultural pest. However, many standard genetic tools are lacking or underdeveloped in this system. Here, we present a set of new reagents to augment existing Tribolium genetic tools. We demonstrate a new GAL4 driver line that employs the promoter of a ribosomal protein gene to drive expression of a UAS responder in the fat body. We also present a novel dual fluorescent reporter that labels cell membranes and nuclei with different fluorophores for the analysis of cellular morphology. This approach also demonstrates the functionality of the viral T2A peptide for bicistronic gene expression in Tribolium. To facilitate classical genetic analysis, we created lines with visible genetic markers by CRISPR-mediated disruption of the yellow and ebony body color loci with a cassette carrying an attP site, enabling future φC31-mediated integration. Together, the reagents presented here will facilitate more robust genetic analysis in Tribolium and serve as a blueprint for the further development of this powerful model's genetic toolkit.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems
  • Gene Expression
  • Gene Order
  • Gene Targeting
  • Genes, Reporter
  • Genetic Techniques*
  • Genetic Vectors / genetics
  • Phenotype
  • Promoter Regions, Genetic
  • Transgenes
  • Tribolium / genetics*

Grants and funding

This work was supported by Indiana University startup funds (to GEZ) and National Science Foundation grant IOS-1353267 (to ACZ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.