Use of Green Fluorescent Protein in mouse embryos.

Green Fluorescent Protein (GFP) has rapidly been established as a versatile and powerful cell marker in many organisms. Initial problems in using it in mammalian cells were solved by introducing mutations to increase its solubility at higher temperatures, such that GFP has now been used as a reporter in both gene expression and cell lineage studies, and to localize proteins within mammalian cells. GFP has two unique advantages: (i) the protein becomes fluorescent in an autocatalytic reaction, so that it can be introduced into any cell type simply as a cDNA or mRNA, or as protein; (ii) it is "bright" enough to be visualized in living cells under conditions that do not cause photodamage to the cells. In this article we outline the ways in which we have used GFP mRNA and cDNA in our studies of mouse cell lineages, and to characterize the behavior of proteins within the embryos.

[1]  M. Ikawa,et al.  A rapid and non‐invasive selection of transgenic embryos before implantation using green fluorescent protein (GFP) , 1995, FEBS letters.

[2]  R Y Tsien,et al.  Understanding, improving and using green fluorescent proteins. , 1995, Trends in biochemical sciences.

[3]  Roger Y. Tsien,et al.  Crystal Structure of the Aequorea victoria Green Fluorescent Protein , 1996, Science.

[4]  M. Zernicka-Goetz,et al.  An indelible lineage marker for Xenopus using a mutated green fluorescent protein. , 1996, Development.

[5]  D. Prasher,et al.  Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Roger Y. Tsien,et al.  Double labelling of subcellular structures with organelle-targeted GFP mutants in vivo , 1996, Current Biology.

[7]  J. Pines,et al.  MPF localization is controlled by nuclear export , 1998, The EMBO journal.

[8]  R Marsault,et al.  Targeting aequorin and green fluorescent protein to intracellular organelles. , 1996, Gene.

[9]  Jim Haseloff,et al.  Mutations that suppress the thermosensitivity of green fluorescent protein , 1996, Current Biology.

[10]  P. Lemaire,et al.  Expression cloning of Siamois, a xenopus homeobox gene expressed in dorsal-vegetal cells of blastulae and able to induce a complete secondary axis , 1995, Cell.

[11]  R Y Tsien,et al.  Wavelength mutations and posttranslational autoxidation of green fluorescent protein. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[12]  R. Tsien,et al.  Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin , 1997, Nature.

[13]  Kenneth R. Spring,et al.  Video Microscopy: The Fundamentals , 1986 .

[14]  S. Kaech,et al.  Application of novel vectors for GFP-tagging of proteins to study microtubule-associated proteins. , 1996, Gene.

[15]  A. Fine,et al.  Live astrocytes visualized by green fluorescent protein in transgenic mice. , 1997, Developmental biology.

[16]  R. Pedersen,et al.  Polarity of the mouse embryo is anticipated before implantation. , 1999, Development.

[17]  S. Rogers,et al.  Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. , 1986, Science.

[18]  B. Hogan,et al.  Manipulating the mouse embryo: A laboratory manual , 1986 .

[19]  Tomoko Nakanishi,et al.  ‘Green mice’ as a source of ubiquitous green cells , 1997, FEBS letters.

[20]  M. Zernicka-Goetz,et al.  Following cell fate in the living mouse embryo. , 1997, Development.