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Generation and characterization of fruitless P1 promoter mutant in Drosophila melanogaster
Journal of Neurogenetics ( IF 1.9 ) Pub Date : 2021-08-02 , DOI: 10.1080/01677063.2021.1931179
Megan C Neville 1 , Alexander Eastwood 1 , Aaron M Allen 1 , Ammerins de Haan 1 , Tetsuya Nojima 1 , Stephen F Goodwin 1
Affiliation  

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 fruΔP1 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.



中文翻译:

黑腹果蝇无果 P1 启动子突变体的产生与表征

摘要

鉴定fruitless ( fru )基因的突变为了解醋蝇Drosophila melanogaster雄性性行为的遗传基础铺平了道路。D. melanogaster雄性对雌性进行精心的求爱展示,最终导致交配。fru的突变已被证明会破坏雄性行为表现的大部分方面,从而使雄性行为不育。水果_基因组位点编码来自多个启动子的多个转录因子亚型;只有受最远端 P1 启动子调控的那些才受性别决定等级的控制,并在雄性特异性行为中发挥作用。在这项研究中,我们使用基于 CRISPR/Cas9 的fru基因靶向基因组编辑来去除 P1 启动子区域。我们已经表明,去除 P1 启动子会导致雄性对雌性的求爱表现和雄性特异性不育显着减少。我们扩大了对fru P1 依赖行为的分析,检查了男性在 12 小时光照:黑暗周期中对求爱歌曲和一般活动水平的反应。我们的新等位基因扩展了可用于fru未来研究的突变库D. melanogaster中的 P1 衍生功能。我们的fru ΔP1突变体将有助于未来研究fru P1 衍生功能,因为它可以纯合而不破坏额外的下游启动子功能,并且可以与其他现存的fru等位基因杂合组合使用。

更新日期:2021-09-21
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