Abstract

The technique of pig cloning holds great promise for the livestock industry, life science, and biomedicine. However, the prenatal death rate of cloned pig embryos is extremely high, resulting in a very low cloning efficiency. This limits the development and application of pig cloning. In this study, we utilized embryo biopsy combined with microproteomics to identify potential factors causing the developmental arrest in cloned pig embryos. We verified the roles of two potential regulators,PDCD6andPLK1, in cloned pig embryo development. We found that siRNA-mediated knockdown ofPDCD6reduced mRNA and protein expression levels of the pro-apoptotic gene,CASP3, in cloned pig embryos.PDCD6knockdown also increased the cleavage rate and blastocyst rate of cloned porcine embryos. Overexpression ofPLK1via mRNA microinjection also improved the cleavage rate of cloned pig embryos. This study provided a new strategy to identify key factors responsible for the developmental defects in cloned pig embryos. It also helped establish new methods to improve pig cloning efficiency, specifically by correcting the expression pattern ofPDCD6andPLK1in cloned pig embryos.Keywords:microproteomics;cloned pig embryos;embryo biopsy;PDCD6;PLK1