Progress and Challenges for Live-cell Imaging of Genomic Loci Using CRISPR-based Platforms

作     者：Xiaotian Wu Shiqi Mao Yachen Ying Christopher J.Krueger Antony K. Chen 

作者机构：Department of Biomedical Engineering College of Engineering Peking University Beijing 100871 China School of Life Sciences Peking University Beijing 100871 China Wallace H Coulter Department of Biomedical Engineering Georgia Institute of Technology Atlanta GA 30332 USA 

出 版 物：《Genomics, Proteomics & Bioinformatics》 (基因组蛋白质组与生物信息学报（英文版）)

年 卷 期：2019年第17卷第2期

页      面：119-128页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 09[农学] 

基　　金：supported by grants from the National Key R&D Program of China (Grant Nos. 2016YFA0501603 and 2016YFA0100702) National Natural Science Foundation of China (Grant No. 31771583) Beijing Natural Science Foundation (Grant No. 7162114) Beijing Municipal R&D Key Project (Grant No. Z151100003915081), China 

主　　题：CRISPR Cas9 dCas9 Genomic imaging sgRNA 

摘      要：Chromatin conformation,localization,and dynamics are crucial regulators of cellular behaviors. Although fluorescence in situ hybridization-based techniques have been widely utilized for investigating chromatin architectures in healthy and diseased states,the requirement for cell fix-ation precludes the comprehensive dynamic analysis necessary to fully understand chromatin activ-ities. This has spurred the development and application of a variety of imaging methodologies for visualizing single chromosomal loci in the native cellular context. In this review,we describe currently-available approaches for imaging single genomic loci in cells,with special focus on clus-tered regularly interspaced short palindromic repeats (CRISPR)-based imaging approaches. In addition,we discuss some of the challenges that limit the application of CRISPR-based genomic imaging approaches,and potential solutions to address these challenges. We anticipate that,with continued refinement of CRISPR-based imaging techniques,significant understanding can be gained to help decipher chromatin activities and their relevance to cellular physiology and pathogenesis.