RNA-seq profiling of the Tg(myo6b:GFP-2A-rpl10a-3xHA) zebrafish hair cell IP vs. whole fish IN

We have developed and tested the efficiency of the Tg(myo6b:GFP-2A-rpl10a-3xHA) zebrafish to specifically enrich for and evaluate the translatome of inner ear and lateral line sensory hair cells (IP) compared to the whole fish transcriptome (IN). We show through RNA-seq that HA-tagged ribosome immunoprecipitation significantly enriches for RNA transcripts of known zebrafish hair cell expressed transcripts. Overall design: Tg(myo6b:GFP-2A-rpl10a-3xHA) were homogenized at 5dpf and the hair cell translatome was enriched for by HA-tagged ribosome immunoprecipitation, followed by RNA extraction. IP samples were compared to input RNA controls (IN) in independent biological triplicates.