Generation and characterization of fruitless P1 promoter mutant in Drosophila melanogaster

Abstract

The identification of mutations in the gene fruitless (fru) paved the way for understanding the genetic basis of male sexual behavior in the vinegar fly Drosophila melanogaster. D. melanogaster males perform an elaborate courtship display to the female, ultimately leading to copulation. Mutations in fru have been shown to disrupt most aspects of the male's behavioral display, rendering males behaviorally sterile. The fru genomic locus encodes for multiple transcription factor isoforms from several promoters; only those under the regulation of the most distal P1 promoter are under the control of the sex determination hierarchy and play a role in male-specific behaviors. In this study, we used CRISPR/Cas9-based targeted genome editing of the fru gene, to remove the P1 promoter region. We have shown that removal of the P1 promoter leads to a dramatic decrease in male courtship displays towards females and male-specific sterility. We have expanded the analysis of fru P1-dependent behaviors, examining male's response to courtship song and general activity levels during12-hour light: dark cycles. Our novel allele expands the mutant repertoire available for future studies of fru P1-derived function in D. melanogaster. Our fru^ΔP1 mutant will be useful for future studies of fru P1-derived function, as it can be homozygosed without disrupting additional downstream promoter function and can be utilized in heterozygous combinations with other extant fru alleles.

Keywords:

• Drosophila
• fruitless
• mutant
• courtship behavior
• CRISPR/Cas9-based targeted genome editing

Acknowledgements

The authors thank Birgit Brüggemeier and Seoho (Michael) Song for support with behavioural assays. We thank Andrew Bassett and Genome Engineering Oxford for analysis of CRISPR reagents. The authors thank current and former members of the Goodwin lab for many helpful discussions over the years.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by a Wellcome Trust Senior Investigator Award to S.F.G [106189/Z/14/Z]. A.E was supported by a Msc Wellcome Trust grant.