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Genome-wide effect of tetracycline, doxycycline and 4-epidoxycycline on gene expression in Saccharomyces cerevisiae.
Yeast ( IF 2.6 ) Pub Date : 2020-07-29 , DOI: 10.1002/yea.3515 Guadalupe Sanchez 1 , Samuel C Linde 1 , Joseph D Coolon 1
Yeast ( IF 2.6 ) Pub Date : 2020-07-29 , DOI: 10.1002/yea.3515 Guadalupe Sanchez 1 , Samuel C Linde 1 , Joseph D Coolon 1
Affiliation
Tetracycline (Tet) and derivative chemicals (e.g., doxycycline or Dox) have gained widespread recognition for their antibiotic properties since their introduction in the late 1970s, but recent work with these chemicals in the lab has shifted to include multiple techniques in all genetic model systems for the precise control of gene expression. The most widely used Tet‐modulated methodology is the Tet‐On/Tet‐Off gene expression system. Tet is generally considered to have effects specific to bacteria; therefore, it should have few off‐target effects when used in eukaryotic systems, and a previous study in the yeast Saccharomyces cerevisiae found that Dox had no effect on genome‐wide gene expression as measured by microarray. In contrast, another study found that the use of Dox in common cell lines and several model organisms led to mitonuclear protein imbalance, suggesting an inhibitory role of Dox in eukaryotic mitochondria. Recently, a new Dox derivative, 4‐epidoxycycline (4‐ED) was developed that was shown to have less off‐target consequences on mitochondrial health. To determine the best tetracycline family chemical to use for gene expression control in S. cerevisiae, we performed RNA sequencing (RNA‐seq) on yeast grown on standard medium compared with growth on media supplemented with Tet, Dox or 4‐ED. We found each caused dozens of genes to change expression, with Dox eliciting the greatest expression responses, suggesting that the specific tetracycline used in experiments should be tailored to the specific gene(s) of interest when using the Tet‐On/Tet‐Off system to reduce the consequences of confounding off‐target responses.
中文翻译:
四环素,强力霉素和4-环氧霉素的全基因组作用对啤酒酵母基因表达的影响。
自从1970年代后期引入四环素(Tet)和衍生化学药品(例如强力霉素或Dox)以来,它们的抗生素特性就得到了广泛认可,但是最近在实验室中使用这些化学药品的工作已经转变为在所有遗传模型系统中包括多种技术用于精确控制基因表达。使用最广泛的Tet调制方法是Tet-On / Tet-Off基因表达系统。Tet通常被认为具有细菌特有的作用;因此,它在真核生物系统中使用时应几乎没有脱靶作用,并且以前在酿酒酵母中已有研究发现通过芯片检测Dox对全基因组基因表达没有影响。相反,另一项研究发现Dox在常见细胞系和几种模型生物中的使用会导致线粒体蛋白失衡,这表明Dox在真核线粒体中具有抑制作用。最近,开发了一种新的Dox衍生物4-环氧丙环素(4-ED),其对线粒体健康的脱靶影响较小。确定用于酿酒酵母基因表达控制的最佳四环素家族化学品,我们对在标准培养基上生长的酵母与在添加了Tet,Dox或4-ED的培养基上生长的酵母进行了RNA测序(RNA-seq)。我们发现每个导致数十个基因改变表达的基因,其中Dox引起最大的表达反应,这表明在使用Tet-On / Tet-Off系统时,实验中使用的特定四环素应针对特定的特定基因进行定制减少混淆脱靶反应的后果。
更新日期:2020-08-24
中文翻译:
四环素,强力霉素和4-环氧霉素的全基因组作用对啤酒酵母基因表达的影响。
自从1970年代后期引入四环素(Tet)和衍生化学药品(例如强力霉素或Dox)以来,它们的抗生素特性就得到了广泛认可,但是最近在实验室中使用这些化学药品的工作已经转变为在所有遗传模型系统中包括多种技术用于精确控制基因表达。使用最广泛的Tet调制方法是Tet-On / Tet-Off基因表达系统。Tet通常被认为具有细菌特有的作用;因此,它在真核生物系统中使用时应几乎没有脱靶作用,并且以前在酿酒酵母中已有研究发现通过芯片检测Dox对全基因组基因表达没有影响。相反,另一项研究发现Dox在常见细胞系和几种模型生物中的使用会导致线粒体蛋白失衡,这表明Dox在真核线粒体中具有抑制作用。最近,开发了一种新的Dox衍生物4-环氧丙环素(4-ED),其对线粒体健康的脱靶影响较小。确定用于酿酒酵母基因表达控制的最佳四环素家族化学品,我们对在标准培养基上生长的酵母与在添加了Tet,Dox或4-ED的培养基上生长的酵母进行了RNA测序(RNA-seq)。我们发现每个导致数十个基因改变表达的基因,其中Dox引起最大的表达反应,这表明在使用Tet-On / Tet-Off系统时,实验中使用的特定四环素应针对特定的特定基因进行定制减少混淆脱靶反应的后果。