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. - Offers cutting edge methods to analyse microRNA expression, function and post-transcriptional modifications - Includes methods to isolate and manipulate cells of the immune system - Provides a description of bioinformatics databases onmiRNAs and their INDICE: 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 inBone Marrow Cells by LNA-FISH.- Measuring microRNA expression in size-limitedF ACS-sorted and microdissected samples.- 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 informalin-fixed paraffin-embedded specimens.- 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.- The 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.- Lentivirus-mediated antagomir expression.- Solution structure ofmiRNA:mRNA complex.- MiRNA editing.- Computational prediction of microRNA targets.- Large-scale integration of microRNA and gene expression data for identification of enriched microRNA:mRNA associations in biological systems.- Identification and validation of the cellular targets of virus-encoded microRNAs.
- ISBN: 978-1-60761-810-2
- Editorial: Humana
- Encuadernacion: Desconocida
- Páginas: 274
- Fecha Publicación: 29/08/2010
- Nº Volúmenes: 1
- Idioma: Inglés