PIWIL3-piRNA Pathway in Rabbit Oogenesis and Early Embryogenesis
We employ RNA Immunoprecipitation sequencing (RIP-seq) to reveal PIWIL3 binding small RNAs as well as mRNAs. overall design: To unravel the molecular mechanisms mediated by PIWIL3, we performed RIP-seq to identify PIWIL3 associated small RNAs and target mRNAs. we used oocytes isolated from wild type rabbit ovaries for PIWIL3 RIP-seq, with IgG RIP-seq as the negative control. We employ smart-seq3 to investigate transcriptome of Piwil3-knockout and WT oocytes/embryos of rabbit. Overall design: In order to investigate endogenous target RNAs of PIWIL3-piRNA complexes, we used Piwil3-Knockout and WT germinal vesicle, germinal vesicle breakdown, meiosis metaphase II stage oocytes, zygote, 2 cell, 4 cell, 8 cell and 16 cell stage embryos to extract total RNAs for single-cell RNA sequencing.