Tunable Protein Stabilization In Vivo Mediated by Shield-1 in Transgenic Medaka

• Published in: PloS one Vol. 10; no. 7; p. e0131252
• Main Authors: Froschauer, Alexander, Kube, Lisa, Kegler, Alexandra, Rieger, Christiane, Gutzeit, Herwig O
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 06.07.2015; Public Library of Science (PLoS)
• Subjects: Animals; Animals, Genetically Modified - metabolism; Caenorhabditis elegans; CRISPR; Developmental biology; Embryos; Fish; Fusion protein; Gene expression; Gene Expression Regulation - drug effects; Genes, Reporter - drug effects; Genetics; Genomes; In vivo methods and tests; Ligands; Male; Morpholines - pharmacology; Organs; Oryzias - genetics; Oryzias - metabolism; Oryzias latipes; Proteasome Endopeptidase Complex - metabolism; Proteasomes; Protein Structure, Tertiary; Proteins; Proteins - metabolism; Reporter gene; Rodents; Spermatogonia; Stem cells; Zebrafish; Zoology
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0131252

Techniques for conditional gene or protein expression are important tools in developmental biology and in the analysis of physiology and disease. On the protein level, the tunable and reversible expression of proteins can be achieved by the fusion of the protein of interest to a destabilizing domain (DD). In the absence of its specific ligand (Shield-1), the protein is degraded by the proteasome. The DD-Shield system has proven to be an excellent tool to regulate the expression of proteins of interests in mammalian systems but has not been applied in teleosts like the medaka. We present the application of the DD-Shield technique in transgenic medaka and show the ubiquitous conditional expression throughout life. Shield-1 administration to the water leads to concentration-dependent induction of a YFP reporter gene in various organs and in spermatogonia at the cellular level.