MicroRNA Protocols

Fully updated and easy to use, this book helps readers explore diverse ways to understand the mechanisms in which miRNAs facilitate the molecular aspects of not only biomedical research but also a wide range of other research fields.

Extensive studies have been conducted on the identification, biogenesis, and processing of microRNA (miRNA) as well as research on the exact mechanism by which miRNAs bring about translational silencing of their targets. In addition, numerous publications point to an important role of miRNAs in development, reprogramming, epigenetics, pathogenesis of cancer, oncogenes and tumor suppressors, biomarkers of various disease onset, and regulation of adipogenesis and obesity; yet many questions still remain. In MicroRNA Protocols, Second Edition, experts in the field provide up-to-date coverage of this diverse area of study. The specific chapters of this edition are related to the analysis of miRNA, targets and expression profiling, various methods to determine its regulation of gene expression, the preparation and isolation of miRNAs in specific tissues, its detection in the saliva, and potential application in cosmetics, wound healing, and prostate cancer. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and known pitfalls.

Fully updated and easy to use, MicroRNA Protocols, Second Edition aims to stimulate readers to explore diverse ways to understand the mechanisms in which miRNAs facilitate the molecular aspects of not only biomedical research but also a wide range of other research fields.

Fully updates and expands upon the successful miRNA methodologies of the first edition Features potential applications in cosmetics, wound healing, and prostate cancer Includes key tips and implementation advice to ensure successful results

Inhalt

The MicroRNA.- Analysis of MicroRNA Length Variety Generated by Recombinant Human Dicer.- A User-Friendly Computational Workflow for the Analysis of miRNA Deep Sequencing Data.- The RNA Gene Information: Retroelement-MicroRNA Entangling as the RNA Quantum Code.- Setting Up an Intronic miRNA Database.- Identify Intronic MicroRNA with Bioinformatics.- Experimental miRNA Target Validation.- Enrichment Analysis of miRNA Targets.- MicroRNA Expression Profiling During Neural Differentiation of Mouse Embryonic Carcinoma P19 Cells.- Evaluating the MicroRNA Targeting Sites by Luciferase Reporter Gene Assay.- Expression Pattern Analysis of MicroRNAs in Caenorhabditis elegans.- Use of Viral Systems to Study miRNA-Mediated Regulation of Gene Expression in Human Cells.- Cloning, Expression, and Functional Analysis of Genomic miRNA Using Retroviral System in Cancer Cells.- Preparing Synaptoneurosomes from Adult Mouse Forebrain.- Isolation of Total RNA and Detection Procedures for miRNA Present in Bovine Cultured Adipocytes and Adipose Tissues.- miRNA and shRNA Expression Vectors Based on mRNA and miRNA Processing.- Gene Silencing In Vitro and In Vivo Using Intronic MicroRNAs.- Vector-Free Methods for Manipulating miRNA Activity In Vitro and In Vivo.- MicroRNA Expression Profiling of Human Induced Pluripotent and Embryonic Stem Cells.- MicroRNA Expression During Neuronal Differentiation of Human Teratocarcinoma NTera2D1 and Mouse Embryonic Carcinoma P19 Cells.- Isolation and Identification of Gene-Specific MicroRNAs.- Transgene-Like Animal Models Using Intronic MicroRNAs.- Mechanism and Method for Generating Tumor-Free iPS Cells Using Intronic MicroRNAmiR302 Induction.- Salivary MicroRNAs and Oral Cancer Detection.- Application of Intronic MicroRNA Agents in Cosmetics.- MicroRNAs in Skin and Wound Healing.- MiRNA Targets of Prostate Cancer.