Real-time functional imaging for monitoring miR-133 during myogenic differentiation

Kato, Yoshio; Miyaki, Shigeru; Yokoyama, Shigetoshi; Omori, Shin; Inoue, Atsushi; Horiuchi, Machiko; Asahara, Hiroshi

doi:10.1016/j.biocel.2009.04.018

Cited by 30 publications

(33 citation statements)

References 31 publications

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“…HEK-293 cells were grown under the same conditions. When C2C12 cells reached a high confluency the medium was changed to differentiation medium comprising DMEM supplemented with 2% HS to induce differentiation (Kato et al, 2009).…”

Section: Reagentsmentioning

confidence: 99%

microRNA133a TargetsFoxl2and Promotes Differentiation of C2C12 into Myogenic Progenitor Cells

Luo

Ling

et al. 2015

DNA and Cell Biology

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microRNAs are endogenous noncoding RNA molecules of *22 nucleotides that regulate gene function by modification of target mRNAs. Due to tissue specific of miR-133a and miR-1/206 for skeletal muscles, we investigated the role of miR-133a and miR-1/206 in promoting the differentiation of the C2C12 cells. The results show that directly transfecting mature miR-133a, miR-1/206, or combinations (miR-1 and miR-206, miR-1 and miR-133a, and miR-133a and miR-206) into C2C12 cells, respectively, for 5 days induces formation of myogenic progenitor cells. Overexpression of miR-133a and miR-206 in C2C12 cells greatly improved multinucleated myotube formation. microRNA-133a (miR-133a) is highly expressed during human muscle development. Using bioinformatics, we identified one putative miR-133a binding site within the 3¢-untranslated region of the mouse Foxl2 mRNA. The expression of Foxl2 was shown to be downregulated by subsequent western blot analysis.

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Section: Reagentsmentioning

confidence: 99%

microRNA133a TargetsFoxl2and Promotes Differentiation of C2C12 into Myogenic Progenitor Cells

Luo

Ling

et al. 2015

DNA and Cell Biology

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“…The high flexibility GFP provides a permanent and heritable label in living cells. By encoding GFP with multitargeting sites of miRNA, lentiviral, or retroviral vectors, in vivo imaging was constructed for viewing [34,35]. Several target sequences for miRNA-142-3p were fused into the 3′-UTR of a GFP expression cassette driven by the ubiquitously expressed phosphoglycerate kinase promoter to develop a lentiviral vector, providing evidence of miRNA regulation in vivo imaging.…”

Section: Mirna Imaging Analysismentioning

confidence: 99%

Intracellular and Organic miRNA In Situ Detection

Zhang

Dong

Tian

2015

SpringerBriefs in Molecular Science

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The detection methods of miRNA in intracellular or organisms fall into two broad categories: indirect detection and direct analysis. The indirect measurement of the expression levels of miRNAs in cells and tissues involves cells lysis and detection by qRT-PCR, northern blotting, or microarray hybridization. The direct analysis methods are a noninvasive manner for repetitively monitoring and obtaining real-time imaging of the intracellular miRNA by using imaging analysis or in situ hybridization (ISH). Technologies for direct detection of the temporal and spatial expression sequence of specific miRNA in cells or tissues are extremely important for elucidating miRNA biology. The progress of optical imaging techniques with multimodal reporter systems holds great promise for noninvasive and real-time imaging of molecular agent expression in living cell. Recent progress in nanotechnology and imaging detection techniques leads to multifunctional nanoprobe with specific-transfection, tracing, and regulation function in intracellular miRNA detection. ISH holds great promise for visualization of the spatial localization of RNA at the tissue, cellular, and even subcellular level.Keywords Intracellular miRNA · Organic miRNA · In situ detection · Cell imaging analysis · Functional nanoprobeThe detection of intracellular miRNA is significant in the development of gene therapy and gene medicines. The deficiency of small size and degradation of the mature miRNAs make it difficult to directly transfer the specific miRNA into the cell. A noninvasive manner for repetitively monitoring and obtaining real-time imaging of the intracellular miRNA is required for the analysis of miRNAs in practical clinic application. Efficient gene vectors, including viral [1] and nonviral categories, [2] were usually required to translocate miRNA probe through membrane barrier into the cell. Retrovirus and adenovirus vectors can be effectively transferred via the gene probes (DNA, RNA) into most cell lines. However, the

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“…A retroviral vector system was developed for elucidating the role of miR-133 in the proliferation of myoblasts during myogenesis [64]. Two FPs of different colors were used to monitor miR-133 activity in living cells, where a GFP reporter was fused to three copies of the target sequence of miR-133 at the 3′UTR, followed by a red fluorescent protein (RFP) gene that is independently expressed under a CMV promoter (not targeted by miR-133).…”

Section: Imaging Mirnas With Fluorescent Protein-based Systemsmentioning

confidence: 99%

“…In the presence of miR-133, GFP expression is suppressed without affecting RFP expression, as evidenced by dual color imaging in differentiated myoblast C2C12 cells. Adapted with permission from [61,64]. …”

Section: Fig (1)mentioning

confidence: 99%

Molecular Imaging Strategies for In Vivo Tracking of MicroRNAs: A Comprehensive Review

Hernandez¹,

Orbay²,

Cai³

2013

CMC

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MicroRNAs (miRNAs) are single-stranded non-coding RNAs of ~22 nucleotides, which can negatively regulate gene expression through induction of mRNA degradation and/or post-transcriptional gene silencing. MiRNAs are key factors in the regulation of many biological processes such as cell proliferation, differentiation, and death. Since miRNAs are known to be in close association with cancer development, non-invasive imaging of miRNA expression and/or activity is of critical importance, for which conventional molecular biology techniques are not suitable or applicable. Over the last several years, various molecular imaging techniques have been investigated for imaging of miRNAs. In this review article, we summarize the current state-of-the-art imaging of miRNAs, which are typically based on fluorescent proteins, bioluminescent enzymes, molecular beacons, and/or various nanoparticles. Non-invasive imaging of miRNA expression and/or biological activity is still at its infancy. Future research on more clinically relevant, non-toxic techniques is required to move the field of miRNA imaging into clinical applications. Non-invasive imaging of miRNA is an invaluable method that can not only significantly advance our understandings of a wide range of human diseases, but also lead to new and more effective treatment strategies for these diseases.

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Real-time functional imaging for monitoring miR-133 during myogenic differentiation

Cited by 30 publications

References 31 publications

microRNA133a TargetsFoxl2and Promotes Differentiation of C2C12 into Myogenic Progenitor Cells

microRNA133a TargetsFoxl2and Promotes Differentiation of C2C12 into Myogenic Progenitor Cells

Intracellular and Organic miRNA In Situ Detection

Molecular Imaging Strategies for In Vivo Tracking of MicroRNAs: A Comprehensive Review

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