Evaluation of FKBP and DHFR based destabilizing domains in Saccharomyces cerevisiae

• Published in: Bioorganic & medicinal chemistry letters Vol. 21; no. 17; pp. 4965 - 4968
• Main Authors: Rakhit, Rishi, Edwards, Sarah R., Iwamoto, Mari, Wandless, Thomas J.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Ltd 01.09.2011; Elsevier
• Subjects: Animals; Biological and medical sciences; chemistry; Conditional gene regulation; Flow Cytometry; Humans; Medical sciences; Mice; mutants; NIH 3T3 Cells; Pharmacology. Drug treatments; Plasmodium; Protein degradation; Saccharomyces cerevisiae; Saccharomyces cerevisiae - growth & development; Saccharomyces cerevisiae - metabolism; Tacrolimus Binding Protein 1A - chemistry; Tacrolimus Binding Protein 1A - metabolism; Tetrahydrofolate Dehydrogenase - chemistry; Tetrahydrofolate Dehydrogenase - metabolism; yeasts; Conditional gene regulation; Protein degradation; Ascomycota; Fungi; Biodegradation; Binding protein; Regulation(control); Enzyme; Oxidoreductases; Dihydrofolate reductase; Saccharomyces cerevisiae
• ISSN: 0960-894X; 1464-3405
• DOI: 10.1016/j.bmcl.2011.06.006

Two orthogonal destabilizing domains have been developed based on mutants of human FKBP12 as well as bacterial DHFR and these engineered domains have been used to control protein concentration in a variety of contexts in vitro and in vivo. FKBP12 based destabilizing domains cannot be rescued in the yeast Saccharomyces cerevisiae; ecDHFR based destabilizing domains are not degraded as efficiently in S. cerevisiae as in mammalian cells or Plasmodium, but provide a starting point for the development of domains with increased signal-to-noise in S. cerevisiae.