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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:
Tanaka, R., Murakami, H., Ote, M., Yamamoto, D.
Source:
Insect molecular biology 2016 v.25 no.4 pp. 355-361
ISSN:
0962-1075
Subject:
Drosophila subobscura, courtship, genes, genetic techniques and protocols, germ cells, mutagenesis, mutants, phenotype, phylogeny, transgenesis
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.
Agid:
5242653