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Bringing MicroRNAs to Light: Methods for MicroRNA Quantification and Visualization in Live Cells

出版社

FRONTIERS MEDIA SA
DOI: 10.3389/fbioe.2020.619583

关键词

microRNA; reporter activity; fluorescence; RT-qPCR; next generation sequencing

资金

  1. Natural Sciences and Engineering Research Council of Canada [RGPIN 04776-2014]
  2. Ontario Ministry of Research and Innovation [ER-18-14-183]
  3. Rare Disease Network [M20-00038]

向作者/读者索取更多资源

MiRNAs are small non-coding RNAs that regulate posttranscriptional gene expression by interacting with target mRNAs. The finely controlled biogenesis, target recognition, and degradation of miRNAs play crucial roles in maintaining cell proliferation, growth, differentiation, and apoptosis. MiRNAs have emerged as potential biomarkers for disease diagnosis, with their expression and activity varying in different contexts such as developmental stages, tissues, and responses to external stimuli.
MiRNAs are small non-coding RNAs that interact with their target mRNAs for posttranscriptional gene regulation. Finely controlled miRNA biogenesis, target recognition and degradation indicate that maintaining miRNA homeostasis is essential for regulating cell proliferation, growth, differentiation and apoptosis. Increasingly, miRNAs have been recognized as a potential biomarker for disease diagnosis. MiRNAs can be found in blood, plasma, and tissues, and miRNA expression and activity differ in developmental stages, tissues and in response to external stimuli. MiRNA transcripts are matured from pri-miRNA over pre-miRNA to mature miRNA, a process that includes multiple steps and enzymes. Many tools are available to identify and quantify specific miRNAs, ranging from measuring total miRNA, specific miRNA activity, miRNA arrays and miRNA localization. The various miRNA assays differ in accuracy, cost, efficiency and convenience of monitoring miRNA dynamics. To acknowledge the significance and increasing research interest in miRNAs, we summarize the traditional as well as novel methods of miRNA quantification with strengths and limitations of various techniques in biochemical and medical research.

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