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揭示微小RNA:活细胞中微小RNA定量与可视化方法

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

作者信息

Siddika Tarana, Heinemann Ilka U

机构信息

Department of Biochemistry, The University of Western Ontario, London, ON, Canada.

出版信息

Front Bioeng Biotechnol. 2021 Jan 18;8:619583. doi: 10.3389/fbioe.2020.619583. eCollection 2020.

Abstract

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.

摘要

微小RNA(miRNA)是一类小的非编码RNA,它们通过与靶mRNA相互作用来进行转录后基因调控。精确调控的miRNA生物合成、靶标识别和降解表明,维持miRNA的稳态对于调节细胞增殖、生长、分化和凋亡至关重要。越来越多的miRNA被认为是疾病诊断的潜在生物标志物。miRNA可存在于血液、血浆和组织中,其表达和活性在发育阶段、组织以及对外界刺激的反应中存在差异。miRNA转录本从初级miRNA(pri-miRNA)经过前体miRNA(pre-miRNA)成熟为成熟miRNA,这一过程涉及多个步骤和多种酶。有许多工具可用于鉴定和定量特定的miRNA,包括测量总miRNA、特定miRNA活性、miRNA阵列以及miRNA定位。各种miRNA检测方法在监测miRNA动态变化的准确性、成本、效率和便利性方面存在差异。为了认识到miRNA的重要性以及对其研究兴趣的不断增加,我们总结了miRNA定量的传统方法和新方法,以及各种技术在生化和医学研究中的优缺点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/470d/7848212/14b4a503c348/fbioe-08-619583-g0001.jpg

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