Department of Medicine, Division of Genetics, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA.
Nat Methods. 2013 Aug;10(8):774-80. doi: 10.1038/nmeth.2558. Epub 2013 Jul 14.
Transcriptional enhancers are a primary mechanism by which tissue-specific gene expression is achieved. Despite the importance of these regulatory elements in development, responses to environmental stresses and disease, testing enhancer activity in animals remains tedious, with a minority of enhancers having been characterized. Here we describe 'enhancer-FACS-seq' (eFS) for highly parallel identification of active, tissue-specific enhancers in Drosophila melanogaster embryos. Analysis of enhancers identified by eFS as being active in mesodermal tissues revealed enriched DNA binding site motifs of known and putative, previously uncharacterized mesodermal transcription factors. Naive Bayes classifiers using transcription factor binding site motifs accurately predicted mesodermal enhancer activity. Application of eFS to other cell types and organisms should accelerate the cataloging of enhancers and understanding how transcriptional regulation is encoded in them.
转录增强子是组织特异性基因表达的主要机制。尽管这些调节元件在发育、对环境压力和疾病的反应中非常重要，但在动物中测试增强子活性仍然很繁琐，只有少数增强子得到了描述。在这里，我们描述了“增强子-FACS-seq”（eFS），用于高度平行地鉴定果蝇胚胎中活跃的、组织特异性的增强子。对 eFS 鉴定为在中胚层组织中活跃的增强子的分析揭示了已知和假定的、以前未被描述的中胚层转录因子的富含 DNA 结合位点基序。使用转录因子结合位点基序的朴素贝叶斯分类器准确地预测了中胚层增强子活性。将 eFS 应用于其他细胞类型和生物体应该会加速增强子的分类，并了解转录调控是如何在其中编码的。
