Pearson Joseph C, McKay Daniel J, Lieb Jason D, Crews Stephen T
Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-3280, USA.
Integrative Program for Biological and Genome Sciences, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-3280, USA.
Development. 2016 Oct 15;143(20):3723-3732. doi: 10.1242/dev.136895.
One of the key issues in studying transcriptional regulation during development is how to employ genome-wide assays that reveals sites of open chromatin and transcription factor binding to efficiently identify biologically relevant genes and enhancers. Analysis of Drosophila CNS midline cell development provides a useful system for studying transcriptional regulation at the genomic level due to a large, well-characterized set of midline-expressed genes and in vivo validated enhancers. In this study, FAIRE-seq on FACS-purified midline cells was performed and the midline FAIRE data were compared with whole-embryo FAIRE data. We find that regions of the genome with a strong midline FAIRE peak and weak whole-embryo FAIRE peak overlap with known midline enhancers and provide a useful predictive tool for enhancer identification. In a complementary analysis, we compared a large dataset of fragments that drive midline expression in vivo with the FAIRE data. Midline enhancer fragments with a midline FAIRE peak tend to be near midline-expressed genes, whereas midline enhancers without a midline FAIRE peak were often distant from midline-expressed genes and unlikely to drive midline transcription in vivo.
发育过程中转录调控研究的关键问题之一是如何利用全基因组分析来揭示开放染色质位点和转录因子结合位点,从而有效地识别生物学相关基因和增强子。果蝇中枢神经系统中线细胞发育的分析为在基因组水平上研究转录调控提供了一个有用的系统,这是因为有大量特征明确的中线表达基因和体内验证的增强子。在本研究中,对通过荧光激活细胞分选(FACS)纯化的中线细胞进行了甲醛辅助的染色质分离测序(FAIRE-seq),并将中线FAIRE数据与全胚胎FAIRE数据进行了比较。我们发现,基因组中具有强烈中线FAIRE峰和微弱全胚胎FAIRE峰的区域与已知的中线增强子重叠,并为增强子识别提供了一个有用的预测工具。在一项补充分析中,我们将一个在体内驱动中线表达的大片段数据集与FAIRE数据进行了比较。具有中线FAIRE峰的中线增强子片段往往靠近中线表达基因,而没有中线FAIRE峰的中线增强子通常远离中线表达基因,并且不太可能在体内驱动中线转录。