NANOS2-binding regulates mRNA half-life and key spermatogonial stem cell self-renewal pathways

We have identified NANOS2 bound targets transcriptome wide by employing CRAC and RNA-seq in mouse SSCs. We have determined the average mRNA half-life in SSCs by SLAM-seq. By comparing CRAC and SLAM-seq datasets, we have demonstrated that NANOS2 binding reduces mRNA stability in SSCs. Overall design: After engineering a Nanos2TAG mouse allele, we derived mouse spermatogonial stem cells (SSCs) from P7 testes. SSCs were expanded in vitro and used to generate libraries for all the sequecing methods described here. First, we have identified NANOS2 RNA occupancy by CRAC, which was normalized by RNA-seq. We then examined global mRNA stability by SLAM-seq and compared this with the relative stability of NANOS2 targets, previously defined by CRAC.