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.
Zernicka-Goetz
M
M
Wellcome/CRC Institute, Tennis Court Road, Cambridge CB2 1QR, United Kingdom. mzg@mole.bio.cam.ac.uk
Pines
J
J
D000818:Animals; D018076:DNA, Complementary / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism*; D017930:Genes, Reporter; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D015335:Molecular Probes; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013696:Temperature
D018076:DNA, Complementary; D008164:Luminescent Proteins; D015335:Molecular Probes; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D012313:RNA