Inhibition of RNAi by Cell Death Signaling

细胞死亡信号传导对 RNAi 的抑制

• 批准号: 0923607
• 负责人: James Birchler
• 金额: $ 30万
• 依托单位: University of Missouri-Columbia
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0923607&HistoricalAwards=false
• 关键词: Inhibition; RNAi; Cell; Death; Signaling

The completion of this project will address unexplored aspects of the RNA interference (RNAi) gene silencing process. In preliminary experiments, it was discovered that cell death produces a signal to adjacent cells that inhibits RNA interference. The research will involve a characterization of modifier mutations that increase or decrease the magnitude of this signaling. The modifier collection will be examined for their effect on the molecular block of RNAi that occurs at the conversion of double stranded RNA to small interfering RNAs. By precise genetic and cytological localization of the mutations recovered, eventually to the single gene level, insight into the molecular basis of the signaling will be revealed by the identification of the corresponding genes involved. Secondly, because cell death signaling increases the expression of retrotransposons, a test of whether novel insertions occur at a greater frequency in conjunction with chronic cell death than in control conditions will be performed.Broader Impacts: Gaining insight in the mechanism of inhibition of RNA interference by cell death signaling will reveal the impact of cell death on viral resistance, transposable element and endogenous gene expression and any consequent impact on mutation rate due to new retrotransposon insertions. RNAi as a reverse genetic tool is impacted by its inhibition by cell death signaling. Thus, an understanding of this process has implications for the basic and applied use of RNAi because its effectiveness could be compromised under such circumstances. The modifier mutation characterization and transposon insertion projects will be conducted by a diverse set of undergraduates so that they will gain a practical appreciation for the principles of classical and modern genetics and potentially be inspired to pursue postgraduate careers. The postdoctoral associate on the project will gain experience in mentoring undergraduates.

该项目的完成将解决 RNA 干扰 (RNAi) 基因沉默过程中尚未探索的方面。在初步实验中，人们发现细胞死亡会向邻近细胞产生抑制RNA干扰的信号。该研究将涉及修饰突变的表征，这些修饰突变会增加或减少这种信号的强度。将检查修饰剂集合对双链 RNA 转化为小干扰 RNA 时发生的 RNAi 分子阻断的影响。通过对恢复的突变进行精确的遗传和细胞学定位，最终达到单基因水平，通过识别所涉及的相应基因，将揭示对信号传导的分子基础的洞察。其次，由于细胞死亡信号传导增加了逆转录转座子的表达，因此将进行一项测试，以确定新的插入是否以比对照条件更高的频率与慢性细胞死亡相关。更广泛的影响：深入了解细胞死亡信号传导抑制RNA干扰的机制将揭示细胞死亡对病毒抗性、转座元件和内源基因表达的影响，以及由于新的插入而对突变率产生的任何后续影响。 逆转录转座子插入。 RNAi 作为一种反向遗传工具，受到细胞死亡信号传导抑制的影响。因此，理解这一过程对于 RNAi 的基本和应用使用具有影响，因为在这种情况下其有效性可能会受到损害。修饰突变表征和转座子插入项目将由不同的本科生进行，以便他们对古典和现代遗传学的原理有实际的了解，并可能受到启发去追求研究生职业。该项目的博士后助理将获得指导本科生的经验。