Miyanari Yusuke
Okazaki Institute for Integrative Bioscience, Okazaki, Japan.
Methods. 2014 Sep;69(2):198-204. doi: 10.1016/j.ymeth.2014.03.030. Epub 2014 Apr 3.
The three-dimensional remodeling of chromatin within nucleus is being recognized as determinant for genome regulation. Recent technological advances in live imaging of chromosome loci begun to explore the biological roles of the movement of the chromatin within the nucleus. To facilitate better understanding of the functional relevance and mechanisms regulating genome architecture, we applied transcription activator-like effector (TALE) technology to visualize endogenous repetitive genomic sequences in mouse cells. The application, called TAL effector-mediated genome visualization (TGV), allows us to label specific repetitive sequences and trace nuclear remodeling in living cells. Using this system, parental origin of chromosomes was specifically traced by distinction of single-nucleotide polymorphisms (SNPs). This review will present our approaches to monitor nuclear dynamics of target sequences and highlights key properties and potential uses of TGV.
细胞核内染色质的三维重塑被认为是基因组调控的决定性因素。染色体位点实时成像技术的最新进展开始探索染色质在细胞核内移动的生物学作用。为了更好地理解调节基因组结构的功能相关性和机制，我们应用转录激活样效应因子（TALE）技术在小鼠细胞中可视化内源性重复基因组序列。这种被称为TAL效应因子介导的基因组可视化（TGV）的应用，使我们能够标记特定的重复序列并追踪活细胞中的核重塑。利用该系统，通过区分单核苷酸多态性（SNP）特异性追踪染色体的亲本来源。本综述将介绍我们监测靶序列核动力学的方法，并重点介绍TGV的关键特性和潜在用途。
