Inhibition of RNAi by Cell Death Signaling

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发出信号，从而抑制RNA干扰。这项研究将涉及修饰突变的特征，这些突变会增加或减少这种信号的大小。将检查修饰剂集合对双链RNA转换为小干扰RNA时发生的RNAi分子块的影响。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，通过识别涉及的相应基因，将揭示信号转导的分子基础。其次，由于细胞死亡信号增加了逆转录转座子的表达，因此将进行一项测试，以确定在慢性细胞死亡的情况下，新插入发生的频率是否高于对照条件。更广泛的影响：深入了解细胞死亡信号抑制RNA干扰的机制将揭示细胞死亡对病毒耐药性、转座元件和内源基因表达的影响，以及新反转录转座子插入对突变率的任何后续影响。RNAi作为一种反向遗传工具，受到细胞死亡信号抑制的影响。因此，对这一过程的理解对RNAi的基本和应用用途具有影响，因为在这种情况下，其有效性可能会受到影响。修饰物突变表征和转座子插入项目将由不同的本科生进行，以便他们将获得对古典和现代遗传学原理的实际欣赏，并可能受到激励，追求研究生职业生涯。该项目的博士后助理将获得指导本科生的经验。

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.