Tyrosinase-Based Reporter Gene for Photoacoustic Imaging of MicroRNA-9 Regulated by DNA Methylation in Living Subjects

MicroRNAs (miRNAs) are a class of negative regulators of gene expression and play critical roles in various biological processes. Conventional approaches for detecting miRNAs, such as northern blotting, microarray, and real-time PCR, usually require the lysis of cell samples and could not provide th...

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Published inMolecular therapy. Nucleic acids Vol. 11; no. C; pp. 34 - 40
Main Authors Zheng, Haifeng, Zhou, Lin, Shi, Yaru, Tian, Jie, Wang, Fu
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.06.2018
Elsevier Limited
American Society of Gene & Cell Therapy
Elsevier
Subjects
Azacytidine
Bioluminescence
Cancer
Cytomegalovirus
Deoxyribonucleic acid
DNA
DNA methylation
DNA microarrays
Enzymes
Gene expression
Laboratory animals
Lysis
Melanin
MicroRNAs
miRNA
miRNAs
Northern blotting
photoacoustic imaging
Reporter gene
tyrosinase
United States > US
miRNAs
tyrosinase
reporter gene
photoacoustic imaging
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Summary:MicroRNAs (miRNAs) are a class of negative regulators of gene expression and play critical roles in various biological processes. Conventional approaches for detecting miRNAs, such as northern blotting, microarray, and real-time PCR, usually require the lysis of cell samples and could not provide the in vivo information about miRNAs in living organisms. Here, we designed a tyrosinase (TYR)-based reporter to monitor miR-9 expression that is regulated by DNA methylation in living cells and animals. During DNA methylation of A549 cells treated by 5-aza-2′-deoxycytidine (5-Aza-dC), the CMV/TYR-3xTS reporter-transfected cells demonstrated a gradual decrease in melanin content, TYR activity, and photoacoustic signal because of the gradual activation of miR-9 expression. The miR-9-regulated repression of TYR activity also resulted in a significant decrease in photoacoustic signal from the flank of mice with 5-Aza-dC treatment, whereas the bioluminescence signal from internal control had no obvious change. The TYR-based miRNA reporter may serve as a new imaging probe for monitoring the dynamic expression of miRNAs during various cellular or disease progression in cells and living animals.
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These authors contributed equally to this work.
ISSN:2162-2531
2162-2531
DOI:10.1016/j.omtn.2018.01.008