Journal
YEAST
Volume 31, Issue 7, Pages 253-264Publisher
WILEY
DOI: 10.1002/yea.3014
Keywords
Schizosaccharomyces pombe; nuclear proteins; anchor-away; rapamycin; FRB; FKBP
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Funding
- National Institutes of Health (NIH) [R01 GM059321, R01 GM081418]
- National Institutes of Health (NIH Cellular, Biochemical, and Molecular Sciences Training Program at the USC Keck School of Medicine)
- National Institutes of Health (USC-Dornsife College Doctoral fellowship)
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Genetic analysis of protein function requires a rapid means of inactivating the gene under study. Typically, this exploits temperature-sensitive mutations or promoter shut-off techniques. We report the adaptation to Schizosaccharomyces pombe of the anchor-away technique, originally designed in budding yeast by Laemmli lab. This method relies on a rapamycin-mediated interaction between the FRB- and FKBP12-binding domains to relocalize nuclear proteins of interest to the cytoplasm. We demonstrate a rapid nuclear depletion of abundant proteins as proof of principle. Copyright (C) 2014 John Wiley & Sons, Ltd.
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