The Drosophila melanogaster attP40 docking site and derivatives are insertion mutations of msp-300.

The Drosophila melanogaster attP40 docking site and derivatives are insertion mutations of msp-300.
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DOI:
10.1371/journal.pone.0278598
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发表时间:
2022
期刊:
影响因子:
3.7
通讯作者:
Stern, Michael
Stern, Michael
中科院分区:
综合性期刊3区
文献类型:
--
作者:
van der Graaf, Kevin;Srivastav, Saurabh;Singh, Pratibha;McNew, James A.;Stern, Michael

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ɸC31 整合酶系统广泛用于黑腹果蝇,以允许转基因靶向特定位点。多年来,已经构建了带有 100 多个 attP 对接位点的果蝇。一个流行的对接位点称为 attP40,位于 Nesprin-1 直系同源物 msp-300 附近,位于某些 msp-300 同工型的上游和其他同工型的第一个内含子内。在这里,我们证明 attP40 会导致幼虫肌肉核聚集,这也是 msp-300 突变赋予的表型。我们还表明,在 attP40 内插入的果蝇在第三龄幼虫中可以表现出 msp-300 转录水平降低。最后,携带某些“转基因 RNAi 项目”(TRiP) 插入 attP40 的染色体可以赋予蛹或成虫无法存活或不育,或在某些遗传背景下产生显性核簇效应。这些表型不需要从 attP40 内的插入进行转录。这些结果表明 attP40 和插入衍生物充当 msp-300 插入突变。在解释来自携带 attP40 的果蝇的数据时应考虑这些发现。
The ɸC31 integrase system is widely used in Drosophila melanogaster to allow transgene targeting to specific loci. Over the years, flies bearing any of more than 100 attP docking sites have been constructed. One popular docking site, termed attP40, is located close to the Nesprin-1 orthologue msp-300 and lies upstream of certain msp-300 isoforms and within the first intron of others. Here we show that attP40 causes larval muscle nuclear clustering, which is a phenotype also conferred by msp-300 mutations. We also show that flies bearing insertions within attP40 can exhibit decreased msp-300 transcript levels in third instar larvae. Finally, chromosomes carrying certain “transgenic RNAi project” (TRiP) insertions into attP40 can confer pupal or adult inviability or infertility, or dominant nuclear clustering effects in certain genetic backgrounds. These phenotypes do not require transcription from the insertions within attP40. These results demonstrate that attP40 and insertion derivatives act as msp-300 insertional mutations. These findings should be considered when interpreting data from attP40-bearing flies.
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