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 Biologya''¢ 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.

MicroRNAs and the Immune System: Methods and Protocols Editor Silvia Monticelli Table of Contents: Table of Contents Preface Contributors Analysis of miRNA expression: ''classic'' methods revisited 1. A rapid, quantitative assay for direct detection of microRNAs and other small RNAs using splinted ligation Sangpen Chamnongpol, Patricia A. Maroney and Timothy W. Nilsen 2. Normalization of microRNA Quantitative RT -PCR Data in Reduced Scale Experimental Designs Gary J. Latham 3. MicroRNA Detection in Bone Marrow Cells by LNA-FISH Silvana Debernardi and Amanda Dixon-McIver 4. Measuring microRNA expression in size-limited F ACS-sorted and microdissected samples Kai P Höfig and Vigo Heissmeyer High-throughput analysis of miRNAs 5. MicroRNA cloning from cells of the immune system Haoquan Wu, Joel Neilson and N. Manjunath 6. High-throughput profiling in the hematopoietic system Muller Fabbri, Riccardo Spizzo and George A. Calin 7. Construction of small RNA cDNA libraries for deep sequencing Molly F. Thomas and K. Mark Ansel 8. microRNA-profiling in formalin-fixed paraffin-embedded specimens Ulrich Lehmann Functional analysis of miRNAs in the immune system: gain-of-function 9. Expression of miRNAs in lymphocytes: a review Raquel Malumbres and Izidore S. Lossos 10. Mouse models for miRNA expression: the ROSA26 locus Stefano Casola 11. Regulation of monocytopoiesis by microRN As Laura Fontana, Antonio Sorrentino and Cesare Peschle 12. MicroRNA activity in B lymphocytes Virginia G. de Yebenes and Almudena R. Ramiro 13. The isolation and characterization of microRNAs of human mature erythrocytes Carolyn Sangokoya, Gregory LaMonte and Jen-Tsan Chi 14. Stable overexpression of miRNAs in bone marrow derived murine mast cells using lentiviral expression vectors Ramon J. Mayoral and Silvia Monticelli 15. Monitoring microRNA activity and validating microRNA targets by reporter-based approaches Alessia Baccarini and Brian D. Brown Functional analysis of miRNAs in the immune system: loss-of-function 16. Lentivirus-mediated antagomir expression Ewa Surdziel, Matthias Eder, and Michaela Scherr miRNA post-transcriptional modifications and mechanisms of action 17. Solution structure ofmiRNA:mRNA complex Mirko Cevec and Janez Plavec 18. MiRNA editing Dylan E. Dupuis and Stefan Maas Bioinformatic analysis and target prediction 19. Computational prediction of microRNA targets Xiaowei Wang 20. Large-scale integration of microRNA and gene expression data for identification of enriched microRNA:mRNA associations in biological systems. Preethi H. Gunaratne, Chad Creighton, Michael Watson and Jayantha B. Tennakoon miRNA and viruses 21. Identification and validation of the cellular targets of virus-encoded microRNAs Kin-Hang Kok, Ting Lei and Dong-Yan Jin