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Clustered regulatory interspaced short palindromic repeats (CRISPR)‐mediated mutagenesis and phenotype rescue by piggyBac transgenesis in a nonmodel Drosophila species
Author(s) -
Tanaka R.,
Murakami H.,
Ote M.,
Yamamoto D.
Publication year - 2016
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/imb.12232
Subject(s) - crispr , biology , drosophila melanogaster , genetics , forward genetics , gene , transgenesis , mutagenesis , transposable element , phenotype , cas9 , genome editing , model organism , mutant , reproductive technology , embryogenesis
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.