Regulation of L1 retrotransposition

描述（由申请人提供）：L1（长穿插核元件1）逆转录转座子是人类基因组中最活跃的可移动元件。它们有超过50万册。L1插入会破坏基因功能，增加基因组的总质量。尽管它们数量众多，但人们对ll是如何调控的知之甚少。在本提案中，我研究了哺乳动物细胞用于抑制基因组L1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方法来监测培养细胞中的L1反转录转位。我们也创造了带有绿色荧光蛋白标记L1转基因的小鼠。更好地了解L1逆转录转位是如何被调控的，将对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.