Profiling Maternal mRNA Translation During Oocyte Development

Abstract

With the progress in our understanding of germ cell development, there is an emerging need to investigate the mechanisms of mRNA translation functioning in these cells. Indeed, posttranscriptional regulations of gene expression drive the most important transitions of the germ cell life cycle. Here we describe a strategy to measure mRNA translation in the oocyte, taking advantage of an approach originally developed to identify the transcriptome of a subgroup of cells in a complex cell mixture. This technique takes advantage of the “RiboTag” approach to express an HA-tag on the large ribosomal subunit of the ribosomes in the oocyte. Immunoprecipitation of the extracts followed by qPCR or RNAseq is used to identify mRNAs actively translated.