dissecting systemic rna interference in the red flour beetle tribolium castaneum parameters affecting the efficiency of rnai解剖系统性rna干扰在红色面粉甲虫种有害castaneum参数影响rnai的效率.pdfVIP

Dissecting Systemic RNA Interference in the Red Flour Beetle Tribolium castaneum: Parameters Affecting the Efficiency of RNAi 1,2 3 1,2 3 Sherry C. Miller , Keita Miyata , Susan J. Brown , Yoshinori Tomoyasu * 1 Division of Biology, Kansas State University, Manhattan, Kansas, United States of America, 2 K-State Arthropod Genomics Center, Kansas State University, Manhattan, Kansas, United States of America, 3 Department of Zoology, Miami University, Oxford, Ohio, United States of America Abstract The phenomenon of RNAi, in which the introduction of dsRNA into a cell triggers the destruction of the corresponding mRNA resulting in a gene silencing effect, is conserved across a wide array of plant and animal phyla. However, the mechanism by which the dsRNA enters a cell, allowing the RNAi effect to occur throughout a multicellular organism (systemic RNAi), has only been studied extensively in certain plants and the nematode Caenorhabditis elegans. In recent years, RNAi has become a popular reverse genetic technique for gene silencing in many organisms. Although many RNAi techniques in non-traditional model organisms rely on the systemic nature of RNAi, little has been done to analyze the parameters required to obtain a robust systemic RNAi response. The data provided here show that the concentration and length of dsRNA have profound effects on the efficacy of the RNAi response both in regard to initial efficiency and duration of the effect in Tribolium castaneum. In addition, our analyses using a series of short dsRNAs and chimeric dsRNA provide evidence that dsRNA cellular uptake (and not the RNAi response itself) is the major step affected by dsRNA size in Tribolium. We also demonstrate that comp