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

doi:10.1021/acschemneuro.8b00614

	Real-Time Functional Bioimaging of Neuron-Specific MicroRNA Dynamics during Neuronal Differentiation Using a Dual Luciferase Reporter
	Zheng, Haifeng 1; Wang, Xinan 1; Chen, Si 1; Shi, Xiaorui 1; Xie, Jinrong 1; Mao, Wenjie 1; Tian, Jie 1,2; Wang, Fu 1
发表期刊	ACS CHEMICAL NEUROSCIENCE
ISSN	1948-7193
	2019-03-01
卷号	10 期号:3 页码:1696-1705
通讯作者	Tian, Jie(jie.tian@ia.ac.cn) ; Wang, Fu(fwang@xidian.edu.cn)
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.
关键词	miRNAs dual-luciferase reporter imaging neurogenesis
DOI	10.1021/acschemneuro.8b00614
关键词[WOS]	GENE
收录类别	SCI
语种	英语
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] ; 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]
项目资助者	National Natural Science Foundation of China ; National Key Research and Development Program of China (973 Program)
WOS研究方向	Biochemistry & Molecular Biology ; Pharmacology & Pharmacy ; Neurosciences & Neurology
WOS类目	Biochemistry & Molecular Biology ; Chemistry, Medicinal ; Neurosciences
WOS记录号	WOS:000462259900074
出版者	AMER CHEMICAL SOC
七大方向——子方向分类	医学影像处理与分析
引用统计	被引频次：3[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.ia.ac.cn/handle/173211/28063
专题	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
通讯作者单位	中国科学院分子影像重点实验室
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.