Abstract
Background
Over the last decade a number of species, from farm animals to rodents, have been cloned using somatic cell nuclear transfer technology (SCNT). This technique has the potential to revolutionize the way that genetically modified animals are made. In its current state, the process of SCNT is very inefficient (<5% success rate), with several technical and biological hurdles hindering development. Yet, SCNT provides investigators with powerful advantages over other approaches, such as allowing for prescreening for the desired level of transgene expression and eliminating the excess production of undesirable wild-type animals. The rat plays a significant role in biomedical research, but SCNT has been problematic for this species. In this study, we address one aspect of the problem by evaluating methods of activation in artificially constructed rat embryos.
Principal Findings
We demonstrate that treatment with a calcium ionophore (ionomycin) combined with a variety of cyclin-dependent kinase inhibitors is an effective way to activate rat embryos. This is in contrast to methods developed for the mouse embryo, which tolerates much less specific chemical treatments. Methods developed to activate mouse embryos do not translate well to rat embryos.
Conclusions
Activation methods developed for one species will not necessarily translate to another species, even if it is closely related. Further, the parthenogenic response to chemical activators is not always a reliable indicator of how reconstructed embryos will react to the same activation method. A better understanding of rat oocyte physiology, although essential for developing better models of disease, may also provide insights that will be useful for making the SCNT process more efficient.
Document Type
Article
Publication Date
3-19-2010
Digital Object Identifier (DOI)
http://dx.doi.org/10.1371/journal.pone.0009799
Repository Citation
Webb, Robin L.; Findlay, Kirk A.; Green, Michael A.; Beckett, Tina L.; and Murphy, M. Paul, "Efficient Activation of Reconstructed Rat Embryos by Cyclin-Dependent Kinase Inhibitors" (2010). Molecular and Cellular Biochemistry Faculty Publications. 4.
https://uknowledge.uky.edu/biochem_facpub/4
Notes/Citation Information
Published in PLoS One, v. 5, no. 3, p. 9799.
© 2010 Webb et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.