Roberts Michael L, Katsoupi Polyxeni, Tseveleki Vivian, Taoufik Era
Synpromics Ltd, 9 Little France Road, Edinburgh, EH16 4UX, UK.
Hellenic Pasteur Institute, Vas Sofias 127, Athens, 11521, Greece.
Methods Mol Biol. 2017;1651:93-112. doi: 10.1007/978-1-4939-7223-4_8.
Synthetic promoters have been developed in a number of different organisms and are capable of mediating specific and enhanced levels of gene expression. Typically, cis-regulatory regions from a few genes are randomly combined to generate a synthetic promoter library, and the sequences with the highest activity are selected for in target cell lines. Here we describe a novel approach that can be employed in the construction of synthetic promoters . Specifically, we use gene expression profiles obtained from microarray datasets to select the cis-regulatory elements that comprise the synthetic promoter library. By adopting this approach, we were able to construct several promoters that could specifically mediate gene expression in colorectal cancer cells. We develop a new selection criteria based on the observed transcriptome of target cells, the frequency that identified cis-regulatory sequences occur in identified gene modules, and the length of identified cis-regulatory regions. Our method allows for the generation of synthetic promoter libraries with increased level of specificity and facilitates the selection of promoters that are highly active only under predefined gene expression profiles.
合成启动子已在多种不同生物体中得到开发，能够介导特定且增强水平的基因表达。通常，从少数基因的顺式调控区域中随机组合以生成合成启动子文库，并在靶细胞系中选择具有最高活性的序列。在此，我们描述了一种可用于构建合成启动子的新方法。具体而言，我们使用从微阵列数据集获得的基因表达谱来选择构成合成启动子文库的顺式调控元件。通过采用这种方法，我们能够构建几个可在结肠癌细胞中特异性介导基因表达的启动子。我们基于观察到的靶细胞转录组、已鉴定的顺式调控序列在已鉴定的基因模块中出现的频率以及已鉴定的顺式调控区域的长度，制定了一种新的选择标准。我们的方法能够生成具有更高特异性水平的合成启动子文库，并有助于选择仅在预定义基因表达谱下具有高活性的启动子。
