The Rsm (Csr) post-transcriptional regulatory system directly controls CRISPR-Cas immunity

RIP-seq analysis to identify RsmA bound RNAs using co-immunoprecipitation and sequencing in Serratia sp. ATCC 39006. We report the binding sites of RsmA (CsrA homologue) in Serratia sp. ATCC39006 Overall design: RsmA was chromosomally tagged using 3xFLAG epitope at the C-terminus in Serratia sp. ATCC39006 wild-type cells and immunoprecipitated using anti-FLAG antibody, untagged version of the wild-type Serratia sp. ATCC39006 strain was used as a control. Lysates for immunoprecipitation for both untagged and RsmA-3xFLAG samples were prepared from 60 O.D. of bacteria grown to mid-log phase ( OD600 ˜ 0.6) . RNA was eluted from both untagged and RsmA-3xFLAG samples to use for cDNA library preparation and sequencing. RNA sequencing of RsmA (NMW7) and LacA strains of stationary phase (12h) cultures for identification of transcriptional changes due to RsmA mutation.