Bystricky Kerstin
University of Toulouse, UPS, Toulouse, France; Laboratoire de Biologie Moléculaire Eucaryote, CNRS, LBME, 31062 Toulouse, France.
FEBS Lett. 2015 Oct 7;589(20 Pt A):3014-22. doi: 10.1016/j.febslet.2015.07.012. Epub 2015 Jul 17.
How chromosomes are folded and how this folding relates to function remain fundamental questions. Answering them is rendered difficult by the stochasticity of chromatin fiber motion which inevitably results in heterogeneity of the populations analyzed. Even if single cell analyses are beginning to yield precious insights, how can we determine whether a snapshot of position is related to function of the probed locus or cell-type? Fluorescence labeling of DNA at single or multiple loci allows determination of their position relative to nuclear landmarks and to each other, enabling us to derive physical parameters of the underlying chromatin fiber. Here I review the contribution of quantitative spatial and temporal analysis of labeled DNA to our understanding of chromosome conformation in different cell types, highlighting live cell imaging techniques and large scale geometrical analysis of multiple loci in 3D.
染色体是如何折叠的,以及这种折叠如何与功能相关,仍然是基本问题。染色质纤维运动的随机性使得回答这些问题变得困难,这种随机性不可避免地导致所分析群体的异质性。即使单细胞分析开始产生宝贵的见解,我们如何确定一个位置的快照是否与被探测位点或细胞类型的功能相关?对单个或多个位点的DNA进行荧光标记,可以确定它们相对于核标记以及彼此之间的位置,使我们能够得出潜在染色质纤维的物理参数。在此,我回顾了对标记DNA进行定量空间和时间分析对我们理解不同细胞类型中染色体构象的贡献,重点介绍了活细胞成像技术和对多个位点进行三维大规模几何分析。