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首页> 外文学位 >RNA interference and dengue virus replication in insect cell culture and Aedes aegypti mosquitoes.
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RNA interference and dengue virus replication in insect cell culture and Aedes aegypti mosquitoes.

机译：昆虫细胞培养和埃及伊蚊中的RNA干扰和登革热病毒复制。

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摘要

RNA interference, or RNAi, can effectively block dengue virus (DEN) replication in mosquito cell culture. The purpose of this study is to examine the mechanism of the RNAi response to DEN-2 virus in mosquito cell culture and to examine the feasibility of RNAi to DEN-2 in the entire mosquito using germ-line transformation. Mosquito cells (C6/36) transformed with a plasmid designed to express double-stranded RNA (dsRNA) derived from the DEN-2 genome do not allow replication of the virus. These cells do not accumulate viral antigen or viral genomic RNA upon infection. The block in DEN-2 replication in these cells was found to be due to an RNAi response, generated by the expression of dsRNA within these cells. The nature of this RNAi response including the dsRNA trigger, the effector molecules (small interfering RNAs or siRNA) and the degradation products of DEN-2 RNA in mosquitoes are defined here. Upon confirmation that the replication silencing observed in mosquito cells was due to RNAi, the process was examined in the mosquito as a whole using a transgenesis approach. Transgenic mosquitoes were engineered to express DEN-2 dsRNA as well as a selectable eyespecific marker. These mosquitoes were tested for their ability to resist DEN-2 infection. None of the transgenic mosquitoes exhibited resistance to DEN-2 infection. The reasons behind the lack of observable DEN-2 resistance in these mosquitoes are discussed along with the pitfalls encountered in the generation of these transgenic mosquitoes.
机译：RNA干扰或RNAi可以有效阻止蚊子细胞培养中的登革热病毒（DEN）复制。这项研究的目的是研究蚊细胞培养中RNAi对DEN-2病毒的应答机制，并使用种系转化技术研究整个蚊子中RNAi对DEN-2病毒的可行性。用表达从DEN-2基因组衍生的双链RNA（dsRNA）的质粒转化的蚊子细胞（C6 / 36）不允许病毒复制。这些细胞在感染后不会积聚病毒抗原或病毒基因组RNA。发现这些细胞中DEN-2复制的阻滞是由于这些细胞中dsRNA表达产生的RNAi反应所致。此处定义了这种RNAi反应的性质，包括dsRNA触发，效应分子（小的干扰RNA或siRNA）和DEN-2 RNA在蚊子中的降解产物。在确认蚊子细胞中观察到的复制沉默是由于RNAi引起的后，使用转基因方法在整个蚊子中检查了这一过程。工程改造了转基因蚊子，以表达DEN-2 dsRNA和选择性眼特异性标记。测试了这些蚊子抵抗DEN-2感染的能力。没有一个转基因蚊子表现出对DEN-2感染的抵抗力。讨论了在这些蚊子中缺乏可观察到的DEN-2抗性的原因，以及在产生这些转基因蚊子时遇到的陷阱。

著录项

  • 作者

    Travanty, Emily Anne.;

  • 作者单位

    Colorado State University.;

  • 授予单位 Colorado State University.;
  • 学科 Biology Microbiology.
  • 学位 Ph.D.
  • 年度 2005
  • 页码 223 p.
  • 总页数 223
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 微生物学;
  • 关键词

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