How genomic shock triggers transposition during hybrid dysgenesis

该项目试图确定可转座元件如何触发基因组不稳定性。转座元件(TES)是自私的DNA元件，可在基因组中增殖并导致突变。60多年前，遗传学家芭芭拉·麦克林托克(Barbara McClintock)证明了DNA损伤可以导致TE激活，但人们对此知之甚少。此外，由于TES本身会造成DNA损伤，一些TES的激活可以触发其他TES的激活。这就是所谓的联合动员。通过将遗传操作与新的基因组测序方法相结合，该项目将确定TES造成的DNA损伤如何触发联合动员。这些结果将为如何在几代人之间保持基因组完整性提供新的见解。该项目将聘请不同的本科生研究人员，提供基因改造和基因组分析方面的培训。此外，它还涉及到向更广泛的社区开发和传播教学工具，包括用于遗传模拟的视频教程和软件模块。为了确定TE动员如何触发其他TE的运动，将以果蝇的杂交发育不良为模型。杂交性不孕是一种不育综合征，是由于父系遗传的TES在生殖系中调节不当而导致的猖獗的易位所致。将进行长读测序，以直接测量世代间的转座和共动员。这一方法将与菌株特异性CRISPR相结合，以确定塑造联合动员概况的遗传因素。关于如何进行联合动员，有两个主要的模型。一种模型认为TE激活是由基因组防御中断引起的。另一种模式是TES由宿主DNA损伤反应的元件直接激活。这两个假设将通过记者分析进行检验。最后，还将使用相同的方法来确定其他模式系统中的联合动员的特征，以确定跨物种的TE联合动员的普遍性。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This project seeks to determine how genome instability is triggered by transposable elements. Transposable elements (TEs) are selfish DNA elements that proliferate through genomes and cause mutation. More than sixty years ago, the geneticist Barbara McClintock demonstrated that DNA damage can lead to TE activation but how this happens is poorly understood. Moreover, since TEs themselves cause DNA damage, the activation of some TEs can trigger the activation of others. This is known as co-mobilization. By combining genetic manipulation with new methods of genome sequencing, this project will determine how the DNA damage caused by TEs triggers co-mobilization. The outcomes will provide new insight into how genome integrity is maintained across generations. This project will engage diverse undergraduate student researchers, providing training in genetic modification and genome analysis. Furthermore, it involves development and dissemination of teaching tools to the broader community, including video tutorials and software modules for genetic simulations.To determine how TE mobilization triggers the movement of other TEs, hybrid dysgenesis in Drosophila virilis will be used as a model. Hybrid dysgenesis is a syndrome of sterility caused by rampant transposition when paternally inherited TEs become mis-regulated in the germline. Long-read sequencing will be performed to directly measure transposition and co-mobilization across generations. This approach will be combined with strain-specific CRISPR to identify genetic factors that shape the profile of co-mobilization. There are two primary models for how co-mobilization may occur. One model proposes that TE activation is caused by disrupted genome defense. An alternate model is that TEs are directly activated by elements of the host DNA damage response. These two hypotheses will be tested using reporter analysis. Finally, the same methods will also be used to characterize co-mobilization in other model systems, in order to determine the generality of TE co-mobilization across species.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.