Anchor Away: A System for Fast Inhibition of Proteins in Drosophila

Methods Mol Biol. 2022:2540:239-249. doi: 10.1007/978-1-0716-2541-5_11.

Abstract

Anchor away is a sequestering method designed to acutely and timely abrogate the function of a protein of interest by anchoring to a cell compartment different from its target. This method induces the binding of the target protein to the anchor by either the addition of rapamycin to Drosophila food or cell media. Rapamycin mediates the formation of a ternary complex between the anchor, which is tagged with the FK506-binding protein (FKBP12), and the target protein fused with the FKB12 rapamycin-binding (FRB) domain of mammalian target of rapamycin (mTOR). The rapamycin-bound target protein stays sequestered away from its compartment, where it cannot perform its biological function.

Keywords: CRISPR; Loss of function; Protein knockout; Rapamycin; TOR.

MeSH terms

  • Animals
  • Drosophila* / metabolism
  • Mammals / metabolism
  • Sirolimus* / pharmacology
  • Tacrolimus Binding Protein 1A / metabolism
  • Tacrolimus Binding Proteins

Substances

  • Tacrolimus Binding Protein 1A
  • Tacrolimus Binding Proteins
  • Sirolimus