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...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-06-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S216225311830009X |
| _version_ | 1819116929181286400 |
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| author | Haifeng Zheng Lin Zhou Yaru Shi Jie Tian Fu Wang |
| author_facet | Haifeng Zheng Lin Zhou Yaru Shi Jie Tian Fu Wang |
| author_sort | Haifeng Zheng |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T05:24:53Z |
| format | Article |
| id | doaj.art-de79fc86c5b148d9ac9f68dac1181835 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-22T05:24:53Z |
| publishDate | 2018-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-de79fc86c5b148d9ac9f68dac11818352022-12-21T18:37:37ZengElsevierMolecular Therapy: Nucleic Acids2162-25312018-06-0111C344010.1016/j.omtn.2018.01.008Tyrosinase-Based Reporter Gene for Photoacoustic Imaging of MicroRNA-9 Regulated by DNA Methylation in Living SubjectsHaifeng Zheng0Lin Zhou1Yaru Shi2Jie Tian3Fu Wang4Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University, Xi’an, Shaanxi 710071, ChinaEngineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University, Xi’an, Shaanxi 710071, ChinaEngineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University, Xi’an, Shaanxi 710071, ChinaEngineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University, Xi’an, Shaanxi 710071, ChinaEngineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University, Xi’an, Shaanxi 710071, ChinaMicroRNAs (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.http://www.sciencedirect.com/science/article/pii/S216225311830009XmiRNAstyrosinasereporter genephotoacoustic imaging |
| spellingShingle | Haifeng Zheng Lin Zhou Yaru Shi Jie Tian Fu Wang Tyrosinase-Based Reporter Gene for Photoacoustic Imaging of MicroRNA-9 Regulated by DNA Methylation in Living Subjects Molecular Therapy: Nucleic Acids miRNAs tyrosinase reporter gene photoacoustic imaging |
| title | Tyrosinase-Based Reporter Gene for Photoacoustic Imaging of MicroRNA-9 Regulated by DNA Methylation in Living Subjects |
| title_full | Tyrosinase-Based Reporter Gene for Photoacoustic Imaging of MicroRNA-9 Regulated by DNA Methylation in Living Subjects |
| title_fullStr | Tyrosinase-Based Reporter Gene for Photoacoustic Imaging of MicroRNA-9 Regulated by DNA Methylation in Living Subjects |
| title_full_unstemmed | Tyrosinase-Based Reporter Gene for Photoacoustic Imaging of MicroRNA-9 Regulated by DNA Methylation in Living Subjects |
| title_short | Tyrosinase-Based Reporter Gene for Photoacoustic Imaging of MicroRNA-9 Regulated by DNA Methylation in Living Subjects |
| title_sort | tyrosinase based reporter gene for photoacoustic imaging of microrna 9 regulated by dna methylation in living subjects |
| topic | miRNAs tyrosinase reporter gene photoacoustic imaging |
| url | http://www.sciencedirect.com/science/article/pii/S216225311830009X |
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