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Anchor Away – A Fast, Reliable and Reversible Technique To Inhibit Proteins in Drosophila melanogaster

Several techniques have been developed to study specific gene function in loss-of-function situations. In Drosophila melanogaster, 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 aboli...

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Autores principales: Bosch, Pablo Sanchez, Pepperl, Julia, Basler, Konrad
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7202031/
https://www.ncbi.nlm.nih.gov/pubmed/32217630
http://dx.doi.org/10.1534/g3.120.401055
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author Bosch, Pablo Sanchez
Pepperl, Julia
Basler, Konrad
author_facet Bosch, Pablo Sanchez
Pepperl, Julia
Basler, Konrad
author_sort Bosch, Pablo Sanchez
collection PubMed
description Several techniques have been developed to study specific gene function in loss-of-function situations. In Drosophila melanogaster, 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 Drosophila, such perdurance is a serious impediment to study gene function. Here we describe the adaptation of the anchor-away technique for use in Drosophila. 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.
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spelling pubmed-72020312020-05-09 Anchor Away – A Fast, Reliable and Reversible Technique To Inhibit Proteins in Drosophila melanogaster Bosch, Pablo Sanchez Pepperl, Julia Basler, Konrad G3 (Bethesda) Investigations Several techniques have been developed to study specific gene function in loss-of-function situations. In Drosophila melanogaster, 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 Drosophila, such perdurance is a serious impediment to study gene function. Here we describe the adaptation of the anchor-away technique for use in Drosophila. 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. Genetics Society of America 2020-03-26 /pmc/articles/PMC7202031/ /pubmed/32217630 http://dx.doi.org/10.1534/g3.120.401055 Text en Copyright © 2020 Sanchez Bosch et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigations
Bosch, Pablo Sanchez
Pepperl, Julia
Basler, Konrad
Anchor Away – A Fast, Reliable and Reversible Technique To Inhibit Proteins in Drosophila melanogaster
title Anchor Away – A Fast, Reliable and Reversible Technique To Inhibit Proteins in Drosophila melanogaster
title_full Anchor Away – A Fast, Reliable and Reversible Technique To Inhibit Proteins in Drosophila melanogaster
title_fullStr Anchor Away – A Fast, Reliable and Reversible Technique To Inhibit Proteins in Drosophila melanogaster
title_full_unstemmed Anchor Away – A Fast, Reliable and Reversible Technique To Inhibit Proteins in Drosophila melanogaster
title_short Anchor Away – A Fast, Reliable and Reversible Technique To Inhibit Proteins in Drosophila melanogaster
title_sort anchor away – a fast, reliable and reversible technique to inhibit proteins in drosophila melanogaster
topic Investigations
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7202031/
https://www.ncbi.nlm.nih.gov/pubmed/32217630
http://dx.doi.org/10.1534/g3.120.401055
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