| 摘要: |
| 基因组的三维空间组织在调节其功能、维持表观遗传和稳定性方面起着重要作用。虽然基于荧光原位杂交的标记技术或单细胞显微观察等方法对此进行了广泛研究，但是细胞内部的时空动态在很大程度上仍难以捉摸。目前标记和观察活细胞中特定内源基因组位点的方法操作难度大且对生物的侵入性伤害较大。CRISPR的发现彻底改变了基因组工程领域。除了基因编辑方面的应用，近年来，CRISPR/Cas9系统在活细胞中特定内源性DNA序列的可视化方面得到一定的发展。通过比较几种不同的成像技术，主要综述基于CRISPR技术在活细胞体内基因组时空成像的基本原理、发展及影响成像效率的主要因素，为探究开发更加简便易行的基于CRISPR成像技术提供一定的思路。 |
| 关键词: 簇状规则间隔的短回文重复 dCas9 sgRNA 活细胞成像 成像效率 |
| DOI：10.14188/j.ajsh.2022.01.003 |
| 分类号:Q522 |
| 基金项目: |
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| Progress on live-cell imaging of genomic loci based on CRISPR/dCas9 technology |
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ZHU Xinghua, WANG Ali, ZHANG Youxuan, LIU Jiangdong
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College of Life Science， Wuhan University， Wuhan 430072，Hubei， China
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| Abstract: |
| The three-dimensional organization of the genome plays an important role in regulating its function， maintaining epigenetic inheritance and stability. Although methods such as fluorescence in situ hybridization-based labeling techniques or single-cell microscopic observations have been extensively studied， the temporal and spatial dynamics inside cells are still largely elusive. The current methods of marking and observing specific endogenous genomic sites in living cells are difficult to operate and are more invasive to organisms. The discovery of CRISPR revolutionizes the field of genome engineering. In addition to the application of gene editing， in recent years， the CRISPR/Cas9 system has made certain developments in the visualization of specific endogenous DNA sequences in living cells. This article compares several different imaging technologies and mainly summarizes the basic principles， development and main factors affecting imaging efficiency of genome spatiotemporal imaging based on CRISPR technology in living cells， and provides certain information for exploring and developing more convenient and feasible CRISPR-based imaging technology. |
| Key words: CRISPR dCas9 sgRNA live cell imaging imaging efficiency |
