Ben Zouari Yousra, Platania Angeliki, Molitor Anne M, Sexton Tom
Institute of Genetics and Molecular and Cellular Biology (IGBMC), Illkirch, France.
CNRS UMR7104, Illkirch, France.
Front Genet. 2020 Jan 21;10:1372. doi: 10.3389/fgene.2019.01372. eCollection 2019.
It is established that transcription of many metazoan genes is regulated by distal regulatory sequences beyond the promoter. Enhancers have been identified at up to megabase distances from their regulated genes, and/or proximal to or within the introns of unregulated genes. The unambiguous identification of the target genes of newly identified regulatory elements can thus be challenging. Well-studied enhancers have been found to come into direct physical proximity with regulated genes, presumably by the formation of chromatin loops. Chromosome conformation capture (3C) derivatives that assess the frequency of proximity between different genetic elements is thus a popular method for exploring gene regulation by distal regulatory elements. For studies of chromatin loops and promoter-enhancer communication, 4C (circular chromosome conformation capture) is one of the methods of choice, optimizing cost (required sequencing depth), throughput, and resolution. For ease of visual inspection of 4C data we present 4See, a versatile and user-friendly browser. 4See allows 4C profiles from the same bait to be flexibly plotted together, allowing biological replicates to either be compared, or pooled for comparisons between different cell types or experimental conditions. 4C profiles can be integrated with gene tracks, linear epigenomic profiles, and annotated regions of interest, such as called significant interactions, allowing rapid data exploration with limited computational resources or bioinformatics expertise.
已确定许多后生动物基因的转录受启动子以外的远端调控序列调节。已在距其调控基因达兆碱基距离处,和/或在非调控基因的内含子近端或内含子内鉴定到增强子。因此,明确鉴定新鉴定的调控元件的靶基因可能具有挑战性。已发现经过充分研究的增强子会与调控基因直接发生物理接近,推测是通过染色质环的形成。因此,评估不同遗传元件之间接近频率的染色体构象捕获(3C)衍生技术是探索远端调控元件对基因调控的常用方法。对于染色质环和启动子-增强子通讯的研究,4C(环状染色体构象捕获)是首选方法之一,它优化了成本(所需的测序深度)、通量和分辨率。为便于直观检查4C数据,我们展示了4See,这是一个通用且用户友好的浏览器。4See允许灵活地将来自同一诱饵的4C图谱绘制在一起,从而可以比较生物学重复样本,或将它们合并以比较不同细胞类型或实验条件。4C图谱可以与基因轨迹、线性表观基因组图谱以及感兴趣的注释区域(如所谓的显著相互作用)整合,从而在有限的计算资源或生物信息学专业知识条件下实现快速的数据探索。