Project description:To investigate whether the regulatory roles of the circadian genes CLOCK and CYCLE as transcription factors in the fruit fly testes and ovaries are similar to those in the head tissue, we conducted ChIP-seq analyses on these three tissues, respectively.
Project description:We used SOLiD technology to deep sequence libraries prepared from 18-35 nt PAGE purfied small RNAs derived from female Drosophila. The objective of our study was to compare the impact of R2D2 or loquacious mutantions on different small RNA populations in the fruit fly.
Project description:Fruit set is triggered after ovule fertilization, as a consequence of the downregulation of ovary growth repressors, such as the tomato transcription factors Auxin/indole-3-acetic acid 9 (IAA9) and Agamous-like 6 (AGL6). We produced small RNA libraries from IAA9- and AGL6-silenced ovaries to identify miRNAs differentially expressed in IAA9- and AGL6-silenced ovaries as compared with unpollinated control ovaries. The identified miRNAs represent a pool of regulatory sRNAs potentially involved in tomato fruit initiation.
Project description:Cucurbits represent an attractive model to explore the dynamics of fruit set. Here we set out to characterize first fruit inhibition (FFI), i.e. the inhibitory effect of the first fruits on subsequent development of younger ovaries during pollination induced fruit set. After the first fertilized ovaries set fruit, younger ovaries fertilized above them on the main stem remained in a temporary state of inhibition. During this stage, the ovaries preserved their size and green color, and if the older fruits were removed within a one-week reversibility window, the inhibited ones set fruit. We compared the gene expression profiles of pollinated ovaries (committed to set fruits) with respect both to those affected by FFI and to non-pollinated ovaries (comitted to senescence). The three fates of the ovaries were characterized at 1 day post anthesis, compare to anthesis, by wide changes in gene expression, with several specific transcripts being up- or down-regulated in response to the presence (or absence) of pollination. Metabolic profiling was undertaken and integrated with the transcriptomic data in order to characterize early physiological changes that occur in post-anthesis ovaries in parthenocarpic and non-parthenocarpic genotypes.
Project description:We sequenced small RNA from total RNA samples in two siRNA-pathway mutants in fly ovaries. This submission comes from a modENCODE project. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf Small RNAs were sequenced from D. melanogaster Canton S ovaries. Raw sequences were clipped by 3' linker sequences recognition, and select clipped sequences longer than 18 nt.
Project description:STING is a protein that plays important role in innate immune response. However, it also has functions not related to immunity. We studied role of STING in fruit fly Drosophila melanogaster. We used microarray to detect gene expression changes in dSTING knockout fruit flies. We studied role of STING in fruit fly Drosophila melanogaster. To detect gene expression changes in dSTING-knockout flies microarray assay was used.
Project description:PIWI-interacting RNAs (piRNAs) are animal gonad-specific small RNAs that control the activity of transposable elements. Long single stranded RNAs from a variety of sources are substrates for the nebulous primary processing pathway that converts these into thousands of 24-30 nucleotide (nt) piRNAs. How these transcripts are selected as precursors is not known. Here we show that targeting a transcript with PIWI slicer activity of cysosolic Ago3 is sufficient to trigger ~30-nt waves of non-overlapping primary piRNAs in the fly ovarian germline. The generated primary piRNAs are almost exclusively loaded into the nuclear PIWI protein, Piwi. In the fly ovarian somatic environment we find that an RNA fragment from the 5? end of a piRNA cluster is able to direct a heterologous sequence into primary processing. This piRNA trigger sequence (PTS) element drives generation of overlapping piRNAs from the transcript. Both mechanisms proceed with general 5?-3? directionality. We propose that the former pathway serves to link cytoplasmic silencing of a target to nuclear transcriptional repression, while the latter extracts silencing information from a wide variety of genomic sources including piRNA clusters, select protein coding and transposon transcripts. Total or immunoprecipitated small RNAs were purified from transfected BmN4 cells, Drosophila ovarian somatic cells (OSC) and from fly ovaries and high-throughput sequencing libraries were prepared. The mouse testicular RNAs were purified after ribozero treatment.
Project description:High-throughput sequencing was used to determine the RNA profiles of the designated genotypes to identify genes with significant expression changes upon deletion of the male germline gene Phf7 in the fruit fly testis.
Project description:Transposable elements are a serious threat for genome integrity and their control via small RNA mediated silencing pathways is an ancient strategy. The fruit fly Drosophila melanogaster has two silencing mechanisms that repress TEs expression: endogenous siRNAs (esiRNAs or endo-siRNAs) and Piwi-interacting small RNAs (piRNAs). The biogenesis of endo-siRNAs involves Loqs-PD, which acts predominantly during processing of dsRNA by Dcr-2, and R2D2 that primarily helps to direct siRNAs for loading into Ago2. We provide deep sequencing evidence consistent with the idea that R2D2 and Loqs-PD can function in part redundantly. Certain transposons display a preference for either dsRBD-protein for production or loading; this appeared to correlate neither with overall abundance, classification of the transposon or a specific site of genomic origin. The endo-siRNA biogenesis pathway in the germline operates according to the same principles as the existing model for the soma, and its impairment does not significantly affect piRNAs. Expanding the analysis, we confirmed the occurrence of somatic piRNA-like RNAs (pilRNAs) that show a ping-pong signature. We detected expression of the Piwi-family protein mRNAs only barely above background, indicating that the somatic pilRNAs may arise from a small sub-population of somatic cells that express a functional piRNA pathway. small RNA sampling experiment; small RNAs were prepared from head & thorax as well as dissected ovaries of Adult female Drosophila melanogaster. We used homozygous mutants of the dsRBD proteins Loqs and r2d2 to determine their contribution to the biogenesis of transposon-derived small RNAs. Heterozygous mutant animals served as control. For each RNA sample, we performed one deep-sequencing run without any treatment, and in parallel one sequencing run after periodate oxidation and beta-elimination. After this treatment, only Ago2, Piwi, Aub and Ago3-loaded small RNAs remain as they carry a 2'-O-methyl modification at their 3'-end. This helps to determine the loading status of the small RNAs detected. In total 8 different RNA samples were prepared and 16 libraries were sequenced.
Project description:The distribtion of H3K18ac in the fruit fly genome was analyzed in wing imaginal discs treated for 2 h with etomoxir and compared to control discs.