Method of Erasing Abnormal Epigenetic Modification in Ovine iCHI Embryo and Use in Generating Gene-Edited Sheep

Abstract

A method of erasing an abnormal epigenetic modification in a sheep iCHI embryo and use thereof. A method including the steps of: transferring a recombinant plasmid expressing protamine into a sheep androgenetic haploid embryonic stem cell, then introducing the cell into a mature sheep oocyte, and performing activation and culture to obtain a sheep iCHI embryo without abnormal epigenetic modifications. In the present disclosure, abnormal methylations in iCHI embryos may be erased by transient expression of protamine in o-haSCs. The blastocyst rate of the prepared Pro-iCHI embryos is significantly higher than that of iCHI embryos, and is similar to that of IVF embryos. The resulting embryos can be used for embryo transfer to obtain semi-cloned gene-edited sheep, providing materials for subsequent breeding applications.

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