Cao Xiangjian, Sun Ying, Lu Peng, Zhao Meiping
Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China.
Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China.
Anal Chim Acta. 2020 Nov 15;1137:225-237. doi: 10.1016/j.aca.2020.08.013. Epub 2020 Aug 23.
Nucleases play crucial roles in maintaining genomic integrity. Visualization of intracellular distribution and translocation of nucleases are of great importance for understanding the in-vivo physiological functions of these enzymes and their roles in DNA repair and other cellular signaling pathways. Here we review the recently developed approaches for fluorescence imaging of nucleases in various eukaryotic cells. We mainly focused on the immunofluorescence techniques, the genetically encoded fluorescent probes and the chemically synthesized fluorescent DNA-substrate probes that enabled in-situ visualization of the subcellular localization of nucleases and their interactions with other protein/DNA molecules within cells. The targeted nucleases included important endonucleases, 3' exonucleases and 5' exonucleases that were involved in the DNA damage repair pathways and the intracellular DNA degradation. The advantages and limitations of the available tools were summarized and discussed.
核酸酶在维持基因组完整性方面发挥着关键作用。可视化核酸酶在细胞内的分布和转位对于理解这些酶的体内生理功能及其在DNA修复和其他细胞信号通路中的作用至关重要。在此，我们综述了最近开发的用于各种真核细胞中核酸酶荧光成像的方法。我们主要关注免疫荧光技术、基因编码荧光探针和化学合成的荧光DNA底物探针，这些方法能够原位可视化核酸酶的亚细胞定位及其与细胞内其他蛋白质/DNA分子的相互作用。靶向核酸酶包括参与DNA损伤修复途径和细胞内DNA降解的重要内切核酸酶、3'外切核酸酶和5'外切核酸酶。我们总结并讨论了现有工具的优缺点。
