Cargando…
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...
Autores principales: | , , |
---|---|
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 |
_version_ | 1783529659593392128 |
---|---|
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. |
format | Online Article Text |
id | pubmed-7202031 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Genetics Society of America |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT boschpablosanchez anchorawayafastreliableandreversibletechniquetoinhibitproteinsindrosophilamelanogaster AT pepperljulia anchorawayafastreliableandreversibletechniquetoinhibitproteinsindrosophilamelanogaster AT baslerkonrad anchorawayafastreliableandreversibletechniquetoinhibitproteinsindrosophilamelanogaster |