Nuclear transfer system for the direct induction of embryonic transcripts from intra- and cross-species nuclei using mouse 4-cell embryos

Junko Tomikawa; Christopher A Penfold; Rena Hatakeyama; Kei Miyamoto

doi:10.1016/j.xpro.2022.101284

Nuclear transfer system for the direct induction of embryonic transcripts from intra- and cross-species nuclei using mouse 4-cell embryos

STAR Protoc. 2022 Apr 11;3(2):101284. doi: 10.1016/j.xpro.2022.101284. eCollection 2022 Jun 17.

Authors

Junko Tomikawa¹, Christopher A Penfold^{2

3

4}, Rena Hatakeyama¹, Kei Miyamoto¹

Affiliations

¹ Laboratory of Molecular Developmental Biology, Faculty of Biology-Oriented Science and Technology, Kindai University, Wakayama 649-6493, Japan.
² Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Site, CB2 3EG Cambridge, UK.
³ Centre for Trophoblast Research, University of Cambridge, Downing Site, CB2 3EG Cambridge, UK.
⁴ Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, Jeffrey Cheah Biomedical Centre, Puddicombe Way, CB2 0AW Cambridge, UK.

Abstract

Reprogramming of somatic nuclei toward the embryonic state has been studied using nuclear transfer (NT) to an oocyte at the metaphase II (MII) stage. In this NT, a somatic nucleus transplanted into an MII oocyte of the same species undergoes DNA replication and cell division before activating embryonic genes. Here, we describe a direct NT protocol using 4-cell stage mouse embryos that enables reprogramming of intra- and cross-species nuclei to express embryonic genes without requiring DNA replication and cell division. For complete details on the use and execution of this protocol, please refer to Tomikawa et al. (2021).

Keywords: Bioinformatics; Cell Biology; Cell culture; Developmental biology; Gene Expression; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Nucleus* / genetics
Cytoplasm
Metaphase
Mice
Nuclear Transfer Techniques*
Oocytes