Abstract
Mammalian oocyte is a highly specialized cell, characterized by synthesis and storage of maternal proteins and RNAs that contributes to the meiotic cell cycle and early embryo development. The fully grown oocyte is transcriptionally quiescent and utilizes only transcripts synthesized and stored during the growing phase. Mouse oocytes are often used as a mammalian model for the study of molecular biology of the cell or biomedical research. Microinjection technique is a useful tool to deliver RNA coding for fluorescently tagged proteins to determine their subcellular localization or function, delivering biosensors for the study of various metabolic pathways or downregulation of specific targets by RNAi or oligo morpholinos to study gene function. Here, we describe a protocol for isolation, cultivation and microinjection of oocytes that might contribute to research or educational purposes.
Keywords: Oocyte, Microinjection, RNA, Meiosis
Materials and Reagents
Equipment
Software
Procedure
Recipes
Acknowledgments
This work was supported by research grant GACR 13-12291S to Andrej Susor. The original work was published in Karabinova et al., (2011) and Susor et al., (2015). Thanks to colleagues from Laboratory of Biochemistry and Molecular Biology of Germ Cells for support.
References
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