Generation and characterization of fruitless P1 promoter mutant in Drosophila melanogaster.

Generation and characterization of fruitless P1 promoter mutant in Drosophila melanogaster.
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黑腹果蝇无果P1启动子突变体的产生及特性

DOI:
10.1080/01677063.2021.1931179
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发表时间:
2021-09
影响因子:
1.9
通讯作者:
Goodwin SF
Goodwin SF
中科院分区:
医学4区
文献类型:
--
作者:
Neville MC;Eastwood A;Allen AM;de Haan A;Nojima T;Goodwin SF

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fruitless(fru)基因突变的鉴定为理解醋蝇果蝇雄性性行为的遗传基础铺平了道路。D.雄性黑腹果蝇向雌性进行精心的求爱展示,最终导致交配。fru基因的突变已经被证明会破坏雄性动物行为表现的大多数方面,使雄性动物行为不育。fru基因组基因座编码来自几个启动子的多种转录因子亚型;只有那些在最远端P1启动子调控下的转录因子亚型受性别决定层次的控制,并在雄性特异性行为中发挥作用。在这项研究中,我们使用了基于CRISPR/Cas9的fru基因靶向基因组编辑,以去除P1启动子区域。我们已经表明,删除P1启动子导致男性求爱显示对女性和男性特定的不育显着减少。我们扩展了对依赖于fru P1的行为的分析,研究了雄性在12小时的光暗周期中对求偶歌的反应和一般活动水平。我们的新等位基因扩展了突变库,可用于未来研究的fru P1衍生功能在D。黑腹菌我们的fru Δ P1突变体将有助于未来fru P1衍生功能的研究,因为它可以在不破坏额外的下游启动子功能的情况下纯合,并且可以与其他现存的fru等位基因进行杂合组合。
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.
DOI: 10.1038/nprot.2009.206
发表时间: 2010-01-01
期刊: NATURE PROTOCOLS
影响因子: 14.8
作者:
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