Lacen Arianna N, Lee Hui-Ting
Department of Chemistry, The University of Alabama at Birmingham, 901 14th Street South, CHEM 274, Birmingham, Alabama 35294-1240, United States.
Chem Biomed Imaging. 2024 Jun 25;2(10):659-682. doi: 10.1021/cbmi.4c00033. eCollection 2024 Oct 28.
Chromatin organization plays a key role in gene regulation throughout the cell cycle. Understanding the dynamics governing the accessibility of chromatin is crucial for insight into mechanisms of gene regulation, DNA replication, and cell division. Extensive research has been done to track chromatin dynamics to explain how cells function and how diseases develop, in the hope of this knowledge leading to future therapeutics utilizing proteins or drugs that modify the accessibility or expression of disease-related genes. Traditional methods for studying the movement of chromatin throughout the cell relied on cross-linking spatially adjacent sections or hybridizing fluorescent probes to chromosomal loci and then constructing dynamic models from the static data collected at different time points. While these traditional methods are fruitful in understanding fundamental aspects of chromatin organization, they are limited by their invasive sample preparation protocols and diffraction-limited microscope resolution. These limitations have been challenged by modern methods based on high- or super-resolution microscopy and specific labeling techniques derived from gene targeting tools. These modern methods are more sensitive and less invasive than traditional methods, therefore allowing researchers to track chromosomal organization, compactness, and even the distance or rate of chromatin domain movement in detail and real time. This review highlights a selection of recently developed methods of chromatin tracking and their applications in fixed and live cells.
染色质组织在整个细胞周期的基因调控中起着关键作用。了解控制染色质可及性的动态过程对于深入了解基因调控、DNA复制和细胞分裂机制至关重要。为了追踪染色质动态以解释细胞如何发挥功能以及疾病如何发展，已经开展了大量研究，希望这些知识能带来未来利用修饰疾病相关基因可及性或表达的蛋白质或药物的治疗方法。传统的研究染色质在整个细胞中运动的方法依赖于交联空间相邻的片段，或将荧光探针与染色体位点杂交，然后根据在不同时间点收集的静态数据构建动态模型。虽然这些传统方法在理解染色质组织的基本方面卓有成效，但它们受到侵入性样本制备方案和衍射极限显微镜分辨率的限制。基于高分辨率或超分辨率显微镜以及源自基因靶向工具的特定标记技术的现代方法对这些限制提出了挑战。这些现代方法比传统方法更灵敏且侵入性更小，因此使研究人员能够详细且实时地追踪染色体组织、紧密程度，甚至染色质结构域的移动距离或速率。本综述重点介绍了最近开发的一些染色质追踪方法及其在固定细胞和活细胞中的应用。
