Project description:To determine functions of circMbl in brain in vivo we generated genetically encoded shRNA directed against individual circRNA-specific back-spliced junctions. We analyzed gene expression by 3' RNA-seq from control (actin-Gal4) and flies expressing the shRNA under the control of a constitutive driver (actin-Gal4; circRNA KD).
Project description:Aim:Transcriptional analysis of the MIN6 beta cell line following control or Paupar lncRNA KD to identify genes regulated by Paupar Methods:MIN6 cells subjected to either control or Paupar KD were transfected for 48 hours before extraction of total RNA using the Qiagen Mini kit. Libraries were prepared from total RNA (RIN>8) with the TruSeq RNA prep kit (Illumina) and sequenced using the HiSeq2000 (Illumina) instrument. More than 30 million reads were mapped to the mouse genome (UCSC/mm9) using Tophat (version 2.0.4) with 4 mismatches and 10 maximum multiple hits. Significantly differentially expressed genes were calculated using DEseq2. Results: We did not observe any signficant changes in genes important for beta cell function Conclusion: Paupar does not have a signficant regulatory function in beta cells
Project description:gene expression profiles in fly brains between wildtype and miR-34 null flies gene expression profiles in fly brains, wild type (3d, 20d) and miR-34 null flies (3d, 20d)
Project description:Flies were fed a normal diet or high-sugar (30% sucrose) diet for 7 days. Brains of flies fasted for 24h or sated were microdissected and immediately frozen in dry ice. The microdissection process from taking the fly out of the vial to freezing the brain took less than a minute. Each brain microdissected piece contains about 50K cells.We dissected 40 brains per condition (4 conditions total) and 4-5 biological replicates