RNA-Seq analysis of MIN6 cells following control KD or Paupar KD

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 Overall design: mRNA profiles of MIN6 cells following either control KD or Paupar KD were generated by deep sequencing, in triplicate, using Illumina HiSeq2000.