ABSTRACT

Until recently, our understanding of chromosome organization in higher eukaryotic cells has been based on analyses of large-scale, low-resolution changes in chromosomes structure. More recently, CRISPR-Cas9 technologies have allowed us to “zoom in” and visualize specific chromosome regions in live cells so that we can begin to examine in detail the dynamics of chromosome organization in individual cells. In this review, we discuss traditional methods of chromosome locus visualization and look at how CRISPR-Cas9 gene-targeting methodologies have helped improve their application. We also describe recent developments of the CRISPR-Cas9 technology that enable visualization of specific chromosome regions without the requirement for complex genetic manipulation.

摘要

直到最近，我们对高等真核细胞中染色体组织的理解一直基于对染色体结构中大规模、低分辨率变化的分析。最近，CRISPR-Cas9 技术使我们能够“放大”并可视化活细胞中的特定染色体区域，以便我们可以开始详细检查单个细胞中染色体组织的动态。在这篇综述中，我们讨论了染色体基因座可视化的传统方法，并研究了 CRISPR-Cas9 基因靶向方法如何帮助改进其应用。我们还描述了 CRISPR-Cas9 技术的最新发展，该技术无需复杂的基因操作即可实现特定染色体区域的可视化。