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 journalArticle

    12 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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