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1 July 2002 Quantitative Analysis of Gene Expression in Preimplantation Mouse Embryos Using Green Fluorescent Protein Reporter
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Abstract

We have developed a method to monitor noninvasively, quantitatively, and in real-time transcription in living preimplantation mouse embryos by measuring expression of a short half-life form of enhanced green fluoresecent protein (EGFP) following microinjection of a plasmid-borne EGFP reporter gene. A standard curve was established by injecting known amounts of recombinant green fluorescent protein, and transcriptional activity was then determined by interpolating the amount of fluoresence in the DNA-injected embryos. This approach permitted multiple measurements in single embryos with no significant detrimental effect on embryonic development as long as light exposure was brief (<30 sec) and no more than two measurements were made each day. This method should facilitate analysis of the regulation of gene expression in preimplantation embryos; in particular, during the maternal-to-zygotic transition, and in other species in which limited numbers of embryos are available.

Serguei Yu Medvedev, Tomoyuki Tokunaga, Richard M. Schultz, Tsutomu Furukawa, Takashi Nagai, Manabu Yamaguchi, Misa Hosoe, Alexander F. Yakovlev, Seiya Takahashi, and Yoshiaki Izaike "Quantitative Analysis of Gene Expression in Preimplantation Mouse Embryos Using Green Fluorescent Protein Reporter," Biology of Reproduction 67(1), 282-286, (1 July 2002). https://doi.org/10.1095/biolreprod67.1.282
Received: 4 December 2001; Accepted: 1 January 2002; Published: 1 July 2002
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