Project description:Purpose: To identify genes that are differentially expressed between control and ski3 mutants Methods: Gene expressions of control and ski3 mutants of third instar larvae were generated by deep sequencing, in two replicates, using Illumina NovaSeq 6000. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Bowtie2 and TopHat followed by Cufflinks. Results: We mapped 800,000-1,200,000 sequence reads per each sample to the fruit fly genome (BDGP6) and identified 93,606 transcripts in the whole body of control and ski3[f03251] with TopHat workflow. Analysis of RNA-seq data idnetified 32 differentially expressed genes relevant to mitochondrial function between control and ski3 mutants with FDR <0.05. The mutation affected over 600 genes, which are involved in many different cellular functions. The results were complex: among 32 affected genes, half of them were upregulated, and the remaining half were downregulated. Six genes were related to the TCA cycle. Of these, the downregulated genes include CG9582 (alpha-ketoglutarate transmembrane transporter), CG32832 (mitochondrial pyruvate carrier), and CG7514 (oxoglutarate:malate antiporter). On the other hand, upregulated genes include AcCoAS (Acetyl-CoA synthase), Sirup (Succinate dehydrogenase), and Men-b (malate dehydrogenase), all of which were directly involved in the TCA cycle. Conclusions: Our study represents the first detailed analysis of ski3 mutant transcriptomes, with biologic replicates, generated by RNA-seq technology. In order to examine the effects of ski3 mutations on gene expression, we performed RNA-seq analyses of wild-type and mutant larvae. Downregulation of transporter, carrier and antiporter suggested a chronic undersupply of TCA cycle substrates, while some of the TCA cycle enzymes were upregulated.
Project description:To investigate how dSETDB1 regulated the genome-wide distribution of HP1 proteins, we performed ChIP-chip assay on the third instar larval lysate from the dSETDB1null mutants in comparison to the wild type. Third instar larvae from the wild type and dSETDB1null mutants were collected. Three independent biological replicates of ChIP with anti-HP1 were performed.
Project description:To investigate how dSETDB1 regulated the genome-wide distribution of HP1 proteins, we performed ChIP-chip assay on the third instar larval lysate from the dSETDB1null mutants in comparison to the wild type.
Project description:We measured gene expression profiles of third instar larvae hemocytes from selected strains related to hemocyte development. Every strain was purchased from commercial companies. And we used microarray data to find what biological processes show abnormalities first, and which gene expressions show similarities and differences between selected strains. Keywords: Strain comparison
Project description:We report here the transcriptomic analysis of Drosophila melanogaster wing imaginal discs from third instar female larvae mutant for corto (cortoL1/corto420) The reference line was the w1118 genetic background of the mutant lines.
Project description:Chronic high sugar feeding induces obesity, hyperglycemia, and insulin resistance in flies and mammals. To gain insight into the mechanisms underlying this response, we profiled gene expression in chronically high sugar fed, wandering (post-prandial) third instar wild type larvae (L3). These data were compared to control-fed larvae as well as those (mid-L3) actively feeding for twelve hours on both diets. We used microarrays to detail the response of Drosophila larvae to high sugar-induced insulin resistance.
Project description:This study describes the epigenetic profiling of the H3K9me2 in wt Drosophila larvae, as well as in Drosophila larvae for which the euchromatic catalytic enzyme depositing H3K9me2 (EHMT) is knocked out. ChIP-Seq profiling of H3K9me2 in wt and EHMT KO third instar Drosophila larvae
Project description:Ectopic expression of DNMT3L in Drosophila causes melanotic tumor in the transgenic flies from fifth generation onwards. We used microaarray data analysis to look for the genes and pathways which are affected on ectopic expression of DNMT3L in Drosophila. Drosophila third instar larvae were selected at particular stages of development for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Our data on third instar larvae tissues of Drosophila melanogaster has allowed us to identify bsAS, a lncRNA antisense to the blistered/DSRF gene, orthologous to the SRF transcription factor in humans, mainly expressed in fly wings. bsAS controls the expression of different isoforms of the blistered gene in Drosophila tissues. CRISPR-CAS deletion of bsAS Transcription Start Site thoroughly abolishes bsAS transcription, inducing a change in the bs isoform usage and the impairment of wing development. Our data consists on stranded RNA-Seq from eye-antenna, leg and wing imaginal discs from third instar larvae. To analyse bsAS mutation, we also performed stranded RNA-Seq of wild type and bsAS-/- eyes and wings from third instar larvae and late pupae.
Project description:We measured gene expression profiles of third instar larvae hemocytes from selected strains related to hemocyte development. Every strain was purchased from commercial companies. And we used microarray data to find what biological processes show abnormalities first, and which gene expressions show similarities and differences between selected strains. Experiment Overall Design: 200~300 Drosophila third instar larvae hemocytes from each of five lines were dissected to get at least 1 microgram of total RNA. Because all samples were hybridized to single-dye Affymetrix chip, control data were compared with data from each mutant.