Anchor away – A fast, reliable and reversible technique to inhibit proteins in Drosophila melanogaster

Abstract

Several techniques have been developed to study specific gene function in loss-of-function situations. In <jats:italic>Drosophila melanoga</jats:italic><jats:italic>st</jats:italic><jats:italic>er</jats:italic>, RNAi and the generation of mutant clones are widely used. However, both techniques have the limitation that there is a significant time lag before gene function is abolished. Given the relatively rapid development of <jats:italic>Drosophila</jats:italic>, such perdurance is a serious impediment to study gene function. Here we describe the adaptation of the anchor-away technique for use in <jats:italic>Drosophila</jats:italic>. Anchor-away was originally developed in yeast to quickly and efficiently abrogate the function of nuclear proteins by sequestering - anchoring - them away in a different cellular compartment. The required components are present in the cells, and the system is triggered by the addition of rapamycin, resulting in a rapid generation of a loss-of-function situation. We provide here proof of principle for the system by producing loss-of-function situations for two nuclear proteins – Pygopus and Brinker. The system allows to study the requirement of any protein during any time window, and at the same time circumvents difficulties, such as off-target effects or variable phenotypes, which are inherent in other techniques, for example RNAi.