Establishment of transgenic lines for jumpstarter method using a composite transposon vector in the ladybird beetle, Harmonia axyridis

PLoS One. 2014 Jun 24;9(6):e100804. doi: 10.1371/journal.pone.0100804. eCollection 2014.

Abstract

In this post-genomic era, genome-wide functional analysis is indispensable. The recent development of RNA interference techniques has enabled researchers to easily analyze gene function even in non-model organisms. On the other hand, little progress has been made in the identification and functional analyses of cis-regulatory elements in non-model organisms. In order to develop experimental platform for identification and analyses of cis-regulatory elements in a non-model organism, in this case, the ladybird beetle, Harmonia axyridis, we established transgenic transposon-tagged lines using a novel composite vector. This vector enables the generation of two types of insertion products (jumpstarter and mutator). The jumpstarter portion carries a transposase gene, while the mutator segment carries a reporter gene for detecting enhancers. The full-composite element is flanked by functional termini (required for movement); however, the mutator region has an extra terminus making it possible for the mutator to remobilize on its own, thus leaving an immobile jumpstarter element behind. Each insertion type is stable on its own, but once crossed, jumpstarters can remobilize mutators. After crossing a jumpstarter and mutator line, all tested G2 females gave rise to at least one new insertion line in the next generation. This jumping rate is equivalent to the P-element-mediated jumpstarter method in Drosophila. These established transgenic lines will offer us the ideal experimental materials for the effective screening and identification of enhancers in this species. In addition, this jumpstarter method has the potential to be as effective in other non-model insect species and thus applicable to any organism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Coleoptera / genetics*
  • DNA Transposable Elements
  • Genetic Engineering / methods*
  • Genetic Vectors
  • Mutagenesis, Site-Directed / methods
  • Organisms, Genetically Modified*
  • Transposases / genetics

Substances

  • DNA Transposable Elements
  • Transposases

Grants and funding

This research was supported in part by Formation and Recognition, PRESTO, Japan Science and Technology Agency, Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan, the Japan Society for the Promotion of Science Research for the Future and Grant-in-Aid (BDP) from the Ministry of Agriculture, Forestry and Fisheries. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.