Clustered regulatory interspaced short palindromic repeats (CRISPR)-mediated mutagenesis and phenotype rescue by piggyBac transgenesis in a nonmodel Drosophila species

R. Tanaka, H. Murakami, M. Ote, D. Yamamoto

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

How behavioural diversity emerged in evolution is an unexplored subject in biology. To tackle this problem, genes and circuits for a behaviour need to be determined in different species for phylogenetic comparisons. The recently developed clustered regulatory interspaced short palindromic repeats/CRISPR associated protein9 (CRISPR/Cas9) system made such a challenge possible by providing the means to induce mutations in a gene of interest in any organism. Aiming at elucidating diversification in genetic and neural networks for courtship behaviour, we attempted to generate a genetic tool kit in Drosophila subobscura, a nonmodel species distantly related to the genetic model Drosophila melanogaster. Here we report the generation of yellow (y) and white mutations with the aid of the CRISPR/Cas9 system, and the rescue of the y mutant phenotype by germline transformation of the newly established y mutant fly line with a y+-marked piggyBac vector. This successful mutagenesis and transformation in D. subobscura open up an avenue for comprehensive genetic analyses of higher functions in this and other nonmodel Drosophila species, representing a key step toward systematic comparisons of genes and circuitries underlying behaviour amongst species.

Original languageEnglish
Pages (from-to)355-361
Number of pages7
JournalInsect Molecular Biology
Volume25
Issue number4
DOIs
Publication statusPublished - 2016 Aug 1

Keywords

  • genome editing
  • germline transformation
  • pigmentation mutations
  • visible marker

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Insect Science

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