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      The anchor-away technique: rapid, conditional establishment of yeast mutant phenotypes.

      1 , ,
      Molecular cell
      Elsevier BV

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          Abstract

          The anchor-away (AA) technique depletes the nucleus of Saccharomyces cerevisiae of a protein of interest (the target) by conditional tethering to an abundant cytoplasmic protein (the anchor) by appropriate gene tagging and rapamycin-dependent heterodimerization. Taking advantage of the massive flow of ribosomal proteins through the nucleus during maturation, a protein of the large subunit was chosen as the anchor. Addition of rapamycin, due to formation of the ternary complex, composed of the anchor, rapamycin, and the target, then results in the rapid depletion of the target from the nucleus. All 43 tested genes displayed on rapamycin plates the expected defective growth phenotype. In addition, when examined functionally, specific mutant phenotypes were obtained within minutes. These are genes involved in protein import, RNA export, transcription, sister chromatid cohesion, and gene silencing. The AA technique is a powerful tool for nuclear biology to dissect the function of individual or gene pairs in synthetic, lethal situations.

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          Author and article information

          Journal
          Mol Cell
          Molecular cell
          Elsevier BV
          1097-4164
          1097-2765
          Sep 26 2008
          : 31
          : 6
          Affiliations
          [1 ] Department of Biochemistry, NCCR Frontiers in Genetics, University of Geneva, 30 Quai Ernest-Ansermet, CH1211-Geneva 4, Switzerland.
          Article
          S1097-2765(08)00540-6
          10.1016/j.molcel.2008.07.020
          18922474
          5f076d99-e7e4-4e41-9783-f4a872d084cf
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