Full Description
In recent years, the critical role of microRNAs has been revealed within the biology of cells that constitute the immune system. In MicroRNAs and the Immune System: Methods and Protocols, expert researchers explore the latest techniques for studying miRNA expression, including the most up-to-date data on splinted ligation and qRT-PCR assays, as well as high-throughput profiling through cloning, deep sequencing, and microarrays. Chapters outline methods to study miRNA functions in various cell types from a single cell type level to entire model organisms, and present studies of miRNAs in the context of viruses and the immune response. Tools are also provided to help navigate bioinformatics databases on miRNAs and their targets. Composed in the highly successful Methods in Molecular Biology™ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.
Contemporary and innovative, MicroRNAs and the Immune System: Methods and Protocols is an essential handbook for immunologists, biochemists, and molecular biologists.
Contents
Analysis of miRNA Expression: Classic Methods Revisited.- A Rapid, Quantitative Assay for Direct Detection of MicroRNAs and Other Small RNAs Using Splinted Ligation.- Normalization of MicroRNA Quantitative RT-PCR Data in Reduced Scale Experimental Designs.- MicroRNA Detection in Bone Marrow Cells by LNA-FISH.- Measuring MicroRNA Expression in Size-Limited FACS-Sorted and Microdissected Samples.- High-Throughput Analysis of miRNAs.- MicroRNA Cloning from Cells of the Immune System.- High-Throughput Profiling in the Hematopoietic System.- Construction of Small RNA cDNA Libraries for Deep Sequencing.- MicroRNA-Profiling in Formalin-Fixed Paraffin-Embedded Specimens.- Functional Analysis of miRNAs in the Immune System: Gain-of-Function.- Expression of miRNAs in Lymphocytes: A Review.- Mouse Models for miRNA Expression: The ROSA26 Locus.- Regulation of Monocytopoiesis by MicroRNAs.- MicroRNA Activity in B Lymphocytes.- Isolation and Characterization of MicroRNAs of Human Mature Erythrocytes.- Stable Overexpression of miRNAs in Bone Marrow-Derived Murine Mast Cells Using Lentiviral Expression Vectors.- Monitoring MicroRNA Activity and Validating MicroRNA Targets by Reporter-Based Approaches.- Functional Analysis of miRNAs in the Immune System: Loss-of-Function.- Lentivirus-Mediated Antagomir Expression.- miRNA Post-Transcriptional Modifications and Mechanisms of Action.- Solution Structure of miRNA:mRNA Complex.- MiRNA Editing.- Bioinformatic Analysis and Target Prediction.- Computational Prediction of MicroRNA Targets.- Large-Scale Integration of MicroRNA and Gene Expression Data for Identification of Enriched MicroRNA-mRNA Associations in Biological Systems.- miRNA and Viruses.- Identification and Validation of the Cellular Targets of Virus-Encoded MicroRNAs.
