Project description:Using quantitative proteomics we identified group of synaptic genes with decreased protein synthesis during homeostatic plasticity. To obtain further information about their mRNA levels/sequences (3’UTR) we performed polyA RNAseq. Using EISA analysis of Ribominus RNAseq dataset we could further differentiate between transcriptional respective post-transcriptional dependent alterations in mRNA levels. In addition Ribominus RNAseq from cell body and processes (dendrites, axons) RNA samples showed us local changes in mRNA levels during homeostatic plasticity. At the end, small RNAseq helped us to identify miRNAs that are increased during homeostatic plasticity and might regulate downregulated genes.
Project description:In order to understand the effect of HHV-6A reactivation on host cell, U2-OS bone osteosarcoma cells were generated carrying latent HHV-6A genome. These cells were either treated with DMSO (solvent control) or Trichostatin-A (TSA) for viral reactivation. As a control cells carrying no HHV-6A were used and treated similarly. Two biological replicates of each sample were processed for small RNA transcriptomics (small RNAseq). Furthermore, HeLa (cervical epithelial cells) were generated carrying lentiviral insertions of one of the small non-coding RNA from HHV-6A (sncRNA-U14). These cells could be transiently induced for sncRNA-U14 transcription using Doxycycline. HeLa cells having a mock lentiviral backbone was used as a control and was treated similarly. Two biological replicates of each sample were processed for small RNA transcriptomics (small RNAseq).