Cargando…

Comparison of GAL80ts and Tet-off GAL80 transgenes

The manipulation of gene expression requires complete control of space, time and level of expression. In Drosophila , the UAS/GAL4 system has been instrumental in achieving spatial control, but the pursuit for the ideal system for temporal control is ongoing. One strategy used by many groups is to u...

Descripción completa

Detalles Bibliográficos
Autores principales: Barwell, Taylor, Geld, Sofia, Seroude, Laurent
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Caltech Library 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10080293/
https://www.ncbi.nlm.nih.gov/pubmed/37033706
http://dx.doi.org/10.17912/micropub.biology.000770
_version_ 1785020890227933184
author Barwell, Taylor
Geld, Sofia
Seroude, Laurent
author_facet Barwell, Taylor
Geld, Sofia
Seroude, Laurent
author_sort Barwell, Taylor
collection PubMed
description The manipulation of gene expression requires complete control of space, time and level of expression. In Drosophila , the UAS/GAL4 system has been instrumental in achieving spatial control, but the pursuit for the ideal system for temporal control is ongoing. One strategy used by many groups is to use GAL80. Two different kinds of GAL80 transgenic constructs have been reported but have never been compared directly. Here, we characterize and compare the repression ability of the GAL80ts and Tet-off GAL80 transgenes by two different assays. We found that GAL80ts was generally more efficient than Tet-off GAL80 and that the level of repression was dependent on the GAL4 driver, and did not necessarily correlate with expression level. We also investigated the level of expression upon inactivation of GAL80ts. The level and kinetics of inactivation were found to differ depending on the GAL4 driver and the stage of the life cycle, and in many cases the inactivation was incomplete. These results emphasize that it is essential to measure experimentally the level of expression under repressed and unrepressed states when multiple GAL4 drivers are used with GAL80 transgenes.
format Online
Article
Text
id pubmed-10080293
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Caltech Library
record_format MEDLINE/PubMed
spelling pubmed-100802932023-04-08 Comparison of GAL80ts and Tet-off GAL80 transgenes Barwell, Taylor Geld, Sofia Seroude, Laurent MicroPubl Biol Materials and Reagents The manipulation of gene expression requires complete control of space, time and level of expression. In Drosophila , the UAS/GAL4 system has been instrumental in achieving spatial control, but the pursuit for the ideal system for temporal control is ongoing. One strategy used by many groups is to use GAL80. Two different kinds of GAL80 transgenic constructs have been reported but have never been compared directly. Here, we characterize and compare the repression ability of the GAL80ts and Tet-off GAL80 transgenes by two different assays. We found that GAL80ts was generally more efficient than Tet-off GAL80 and that the level of repression was dependent on the GAL4 driver, and did not necessarily correlate with expression level. We also investigated the level of expression upon inactivation of GAL80ts. The level and kinetics of inactivation were found to differ depending on the GAL4 driver and the stage of the life cycle, and in many cases the inactivation was incomplete. These results emphasize that it is essential to measure experimentally the level of expression under repressed and unrepressed states when multiple GAL4 drivers are used with GAL80 transgenes. Caltech Library 2023-03-23 /pmc/articles/PMC10080293/ /pubmed/37033706 http://dx.doi.org/10.17912/micropub.biology.000770 Text en Copyright: © 2023 by the authors https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Materials and Reagents
Barwell, Taylor
Geld, Sofia
Seroude, Laurent
Comparison of GAL80ts and Tet-off GAL80 transgenes
title Comparison of GAL80ts and Tet-off GAL80 transgenes
title_full Comparison of GAL80ts and Tet-off GAL80 transgenes
title_fullStr Comparison of GAL80ts and Tet-off GAL80 transgenes
title_full_unstemmed Comparison of GAL80ts and Tet-off GAL80 transgenes
title_short Comparison of GAL80ts and Tet-off GAL80 transgenes
title_sort comparison of gal80ts and tet-off gal80 transgenes
topic Materials and Reagents
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10080293/
https://www.ncbi.nlm.nih.gov/pubmed/37033706
http://dx.doi.org/10.17912/micropub.biology.000770
work_keys_str_mv AT barwelltaylor comparisonofgal80tsandtetoffgal80transgenes
AT geldsofia comparisonofgal80tsandtetoffgal80transgenes
AT seroudelaurent comparisonofgal80tsandtetoffgal80transgenes