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Real-Time Functional Bioimaging of Neuron-Specific MicroRNA Dynamics during Neuronal Differentiation Using a Dual Luciferase Reporter

文献类型：期刊论文

作者Zheng, Haifeng1; Wang, Xinan1; Chen, Si1; Shi, Xiaorui1; Xie, Jinrong1; Mao, Wenjie1; Tian, Jie1,2; Wang, Fu1
刊名ACS CHEMICAL NEUROSCIENCE
出版日期2019-03-01
卷号10期号:3页码:1696-1705
关键词miRNAs dual-luciferase reporter imaging neurogenesis
ISSN号1948-7193
DOI10.1021/acschemneuro.8b00614
通讯作者Tian, Jie(jie.tian@ia.ac.cn) ; Wang, Fu(fwang@xidian.edu.cn)
英文摘要The capability of monitoring the neuronal differentiation process in living cells is crucial to the understanding of neuronal development and the practical application of cell therapy for the treatment of neurodegenerative disorders. Current research methods, including Northern blot and real-time PCR analysis, have been extensively employed to quantify miRNA expression during cellular processes. However, these methods require cell destruction and could not provide dynamic information on miRNA expression and function in living organisms. In the present study, we developed a dual luciferase reporter to monitor the expression pattern of neuron-specific miRNA-9 and miRNA-124a during neuronal differentiation in vitro and in vivo. The miRNA-responsive reporter was designed to encode a firefly luciferase (Fluc) gene containing miRNA target sequences and a Renilla luciferase (Rluc) gene for normalization. These two genes were independent modules and transcribed by two different promoters, which enables precise sensing miRNA activity without mutual transcription interference. We demonstrated that the functional activation of miRNA-9 and miRNA-124a during neurogenesis was visualized by the reduction of Fluc bioluminescence signal in P19 cells and nude mice without Rluc signal change, suggesting that miRNA-9 and miRNA-124a specifically downregulated their targets in accordance with their expression. Our dual luciferase-based miRNA imaging system provides a useful tool to quantitatively and continuously monitor miRNA activity during various biological processes.
WOS关键词GENE
资助项目National Natural Science Foundation of China[81571721] ; National Natural Science Foundation of China[81772010] ; National Key Research and Development Program of China (973 Program)[2017YFA0205202]
WOS研究方向Biochemistry & Molecular Biology ; Pharmacology & Pharmacy ; Neurosciences & Neurology
语种英语
WOS记录号WOS:000462259900074
出版者AMER CHEMICAL SOC
资助机构National Natural Science Foundation of China ; National Key Research and Development Program of China (973 Program)
源URL[http://ir.ia.ac.cn/handle/173211/28062]  
专题自动化研究所_中国科学院分子影像重点实验室
通讯作者Tian, Jie; Wang, Fu
作者单位1.Xidian Univ, Sch Life Sci & Technol, Minist Educ, Engn Res Ctr Mol & Neuro Imaging, Xian 710071, Shaanxi, Peoples R China
2.Chinese Acad Sci, Inst Automat, CAS Key Lab Mol Imaging, Beijing 100190, Peoples R China
推荐引用方式
GB/T 7714		 Zheng, Haifeng,Wang, Xinan,Chen, Si,et al. Real-Time Functional Bioimaging of Neuron-Specific MicroRNA Dynamics during Neuronal Differentiation Using a Dual Luciferase Reporter[J]. ACS CHEMICAL NEUROSCIENCE,2019,10(3):1696-1705.
APA Zheng, Haifeng.,Wang, Xinan.,Chen, Si.,Shi, Xiaorui.,Xie, Jinrong.,...&Wang, Fu.(2019).Real-Time Functional Bioimaging of Neuron-Specific MicroRNA Dynamics during Neuronal Differentiation Using a Dual Luciferase Reporter.ACS CHEMICAL NEUROSCIENCE,10(3),1696-1705.
MLA Zheng, Haifeng,et al."Real-Time Functional Bioimaging of Neuron-Specific MicroRNA Dynamics during Neuronal Differentiation Using a Dual Luciferase Reporter".ACS CHEMICAL NEUROSCIENCE 10.3(2019):1696-1705.

入库方式： OAI收割

来源：自动化研究所

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