Functional Analysis of the Drosophila Dnop5 Using Targeted RNA Interference
نویسندگان
چکیده
Dnop5 is a member of the conserved nop5/sik1 gene family, which encode components of small nucleolar ribonucleoprotein(snoRNP) complexes. To study the function of DNop5, we generated the polyclonal antibody and determined its expression pattern. It is highly expressed in different periods of the Drosophila development. We used heritable RNA interference (RNAi) in combination with the yeast GAL4/UAS binary system to knock down the DNop5 protein. It resulted in lethality and dramatic somatic anomalies in RNAi mutant fly, in which the DNop5 protein is reduced efficiently. Northern blotting showed that the processing of 18S rRNA was disrupted in DNop5 knock down fly, but 28S rRNA is normal. These results suggest that DNop5 is essential for the Drosophila growth and function in the execution of early pre-rRNA processing steps that lead to formation of 18S rRNA.
منابع مشابه
Restricted expression and subnuclear localization of the Drosophila gene Dnop5, a member of the Nop/Sik family of the conserved rRNA processing factors
Members of the conserved nop5/sik1 gene family encode components of small nucleolar ribonucleoprotein (snoRNP) complexes, which have an essential function in rRNA-processing. We describe a novel Drosophila member of this family, termed Dnop5. The gene is expressed in nurse cells during oogenesis and transcripts are deposited into the growing oocyte. Maternal transcripts become evenly distribute...
متن کاملBcr-abl Silencing by Specific Small-Interference RNA Expression Vector as a Potential Treatment for Chronic Myeloid Leukemia
Background: RNA interference (RNAi) is the mechanism of gene silencing-mediated messenger RNA degradation by small interference RNA (siRNA), which becomes a powerful tool for in vivo research, especially in the areas of cancer. In this research, the potential use of an expression vector as a specific siRNA producing tool for silencing of Bcr-abl in K562 cell line has been investigated. Methods:...
متن کاملInjection of dsRNA into Drosophila Embryos for RNA Interference (RNAi).
INTRODUCTIONRNA interference (RNAi) is a powerful method for determining the role of specific genes during Drosophila embryogenesis. This protocol describes a technique by which Drosophila embryos can be injected with dsRNA in order to disrupt targeted gene function. The approach is straightforward, utilizing improved methods for injecting the dsRNA directly through the chorion of the embryo. T...
متن کاملReagent and Data Resources for Investigation of RNA Binding Protein Functions in Drosophila melanogaster Cultured Cells
RNA binding proteins (RBPs) are involved in many cellular functions. To facilitate functional characterization of RBPs, we generated an RNA interference (RNAi) library for Drosophila cell-based screens comprising reagents targeting known or putative RBPs. To test the quality of the library and provide a baseline analysis of the effects of the RNAi reagents on viability, we screened the library ...
متن کاملRNAi-induced off-target effects in Drosophila melanogaster: frequencies and solutions.
Genes can be silenced with short-interfering RNA molecules (siRNA). siRNAs are widely used to identify gene functions and have high potential for therapeutic treatments. It is critical that the siRNA specifically targets the expression of the gene of interest but has no off-target effects on other genes. Although siRNAs were initially considered to be exclusively active on mature mRNAs in the c...
متن کامل