Barwell Taylor, Geld Sofia, Seroude Laurent
Biology, Queen's University, Kingston, Ontario, Canada.
MicroPubl Biol. 2023 Mar 23;2023. doi: 10.17912/micropub.biology.000770. eCollection 2023.
The manipulation of gene expression requires complete control of space, time and level of expression. In , 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.
基因表达的操控需要对表达的空间、时间和水平进行完全控制。在[具体情况未提及]中，UAS/GAL4系统在实现空间控制方面发挥了重要作用，但对理想的时间控制体系的探索仍在继续。许多研究小组采用的一种策略是使用GAL80。已报道了两种不同类型的GAL80转基因构建体，但从未直接进行过比较。在这里，我们通过两种不同的检测方法对GAL80ts和Tet-off GAL80转基因的抑制能力进行了表征和比较。我们发现，GAL80ts通常比Tet-off GAL80更有效，并且抑制水平取决于GAL4驱动子，不一定与表达水平相关。我们还研究了GAL80ts失活后的表达水平。发现失活的水平和动力学因GAL4驱动子和生命周期阶段而异，并且在许多情况下失活并不完全。这些结果强调，当多个GAL4驱动子与GAL80转基因一起使用时，通过实验测量抑制和未抑制状态下的表达水平至关重要。
