Kiefer Christine M, Dean Ann
Laboratory of Cellular and Developmental Biology, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2012;833:29-45. doi: 10.1007/978-1-61779-477-3_3.
In metazoans transcriptional enhancers and their more complex relatives, locus control regions, are often located at great linear distances from their target genes. In addition, these elements frequently activate different members of gene families in temporal sequence or in different tissues. These issues have complicated understanding the mechanisms underlying long-range gene activation. Advances in primarily technical approaches, such as chromosome conformation capture (3C) and its derivatives have now solidified the idea that distant regulatory elements achieve proximity with their target genes when they are activating them. Furthermore, these approaches are now allowing genome-wide views of chromosome interactions that are likely to include regulatory, structural, and organization aspects from which we will be able to understand more about nuclear structure. At the base of these advances are experimental approaches to localize protein-binding sites in chromatin, to assess remodeling of chromatin and to measure interaction frequency between distant sites. Examples of these approaches comprise this review.
在后生动物中,转录增强子及其更复杂的相关元件——基因座控制区,通常与其靶基因在直线距离上相隔很远。此外,这些元件常常按时间顺序或在不同组织中激活基因家族的不同成员。这些问题使得理解远程基因激活背后的机制变得复杂。诸如染色体构象捕获(3C)及其衍生技术等主要技术方法的进展,现已巩固了这样一种观点,即远距离调控元件在激活靶基因时会与它们靠近。此外,这些方法现在能够对全基因组范围的染色体相互作用进行观察,这些相互作用可能包括调控、结构和组织等方面,从中我们将能够更多地了解核结构。这些进展的基础是用于在染色质中定位蛋白质结合位点、评估染色质重塑以及测量远距离位点之间相互作用频率的实验方法。这些方法的实例构成了本综述的内容。