Regulation of L1 retrotransposition

描述（由申请人提供）：L1（长分散核元件1）反转录转座子是人类基因组中最活跃的移动的元件。目前已发行50多万册。L1插入可以破坏基因功能并增加基因组的整体质量。尽管它们丰富，但人们对LLS是如何调节的知之甚少。在这个提议中，我调查了哺乳动物细胞用于包含L1对基因组的损伤的三种潜在策略：1。修复由L1插入引起或用于L1插入的双链DNA断裂。2.通过凋亡杀死具有破坏性L1插入的细胞。3.尝试通过使用RNA干扰破坏L1 RNA来阻止L1插入。为了研究这些假设的L1调节模式，我的实验室开发了一种流式细胞术检测培养细胞中L1逆转录转座。我们还创造了带有绿色荧光蛋白标记的L1转基因的小鼠。更好地了解L1逆转录转座是如何调节的，应该对L1生物学、机制和L1在癌症中促进基因组不稳定性的潜力产生重要的见解。

DESCRIPTION (provided by applicant): L1 (Long Interspersed Nuclear Element 1) retrotransposons are the most active mobile elements in the human genome. They are present in over 500,000 copies. L1 insertions can disrupt gene function and add to the overall mass of the genome. Despite their abundance, very little is known about how Lls are regulated. In this proposal, I investigate three potential strategies used by mammalian cells to contain L1 damage to the genome: 1. Repair double stranded DNA breaks that are caused by or used for L1 insertions. 2. Kill cells with damaging L1 insertions by apoptosis. 3. Try to prevent L1 insertion by using RNA interference to destroy L1 RNA. To investigate these hypothesized modes of L1 regulation, my lab has developed a flow cytometric assay to monitor L1 retrotransposition in cultured cells. We have also created mice with green fluorescent protein tagged L1 transgenes. A better understanding of how L1 retrotransposition is regulated should yield important insights into L1 biology, mechanism and the potential of L1s to contribute to genomic instability in cancer.