Identification of MBL-1 target transcript through RIP-seq.
MBL-1 is an RNA-binding protein important for the differentiation and development of many animal tissues. Our aim was to identify the binding targets of the MBL-1 protein in C. elegans. RNA from a CRISPR/CAS9-generated strain expressing a 3xFLAG-tagged MBL-1 protein was immunopreciprecipitated with an anti-FLAG antibody. RNA-seq was then performed to identify binding targets of the RNA-binding protein MBL-1. Experiments were performed at the L4 stage. Overall design: RNA samples were collected from L4 stage 3xFLAG::MBL-1 animlas before (RNA input) and after immunoprecipitation with an anti-FLAG antibody (IP samples). Three replicates for each sample were made. Following RNA-seq, differentially expressed peaks (and thus MBL-1 binding targets) were identified by comparison of input and IP sample peaks.