Bulk-RNA sequencing of E8.25 posterior tissue +2 somites from controls and Sp5/8 double knockout or Wnt3a knockout embryos
Sp5/8 are Zn finger downstream transcription factors of Wnt/b-catenin signaling pathway.  Deletion of Sp5/8 in mice result in severer posterior axis truncation by E9.5 closely resembling Wnt pathway mutants.  The goal of this project is to identify differentially expressed genes between controls and Sp5/8 double knockout mutants that could potentially reveal mechanisms of the role of Sp5/8 factors in self-renewal and differentiation of caudal progenitors. Overall design: Posterior tissue + 2 somites were dissected out from E8.25 embryos (3-5 somites).  Total RNA was isolated using Trizol reagent according to the manufacturer's instructions (ThermoFisher Scientific) and genomic DNA was eliminated using Turbo-DNAse.  RNA quality was assessed using Agilent RNA bioanalyzer and samples with RNA integrity numbers greater than 9.0 were selected for sequencing.