MicroRNAs and the immune system : methods and protocols / edited by Silvia Monticelli. — New York, NY : Humana Press, c2010. – (58.17/M592/v.667)

Contents

    CONTENTS
    
    Preface v
    Acknowledgments vii
    Contributors xi
    PART I ANALYSIS OF MIRNA EXPRESSION" CLASSIC METHODS REVISITED
    1. A Rapid， Quantitative Assay for Direct Detection of MicroRNAs and Other Small RNAs Using Splinted Ligation 3
    2. Normalization of MicroRNA Quantitative RT-PCR Data in Reduced Scale Experimental Designs 19
    3. MicroRNA Detection in Bone Marrow Cells by LNA-FISH
    4. Measuring MicroRNA Expression in Size-Limited FACS-Sorted and Microdissected Samples 47
    PART II HIGH-THROUGHPUT ANALYSIS OF miRNAs
    5. MicroRNA Cloning from Cells of the Immune System
    6. High-Throughput Profiling in the Hematopoietic System
    7. Construction of Small RNA cDNA Libraries for Deep Sequencing 93
    8. MicroRNA-Profiling in Formalin-Fixed Paraffin-Embedded Specimens 113
    PART III FUNCTIONAL ANALYSIS OF miRNAs IN THE IMMUNE SYSTEM: GAIN-OF-FUNCTION
    9. Expression of miRNAs in Lymphocytes: A Review
    10. Mouse Models for miRNA Expression: The ROSA26 Locus 145
    11. Regulation of Monocytopoiesis by MicroRNAs 165
    12. MicroRNA Activity in B Lymphocytes 177
    13. Isolation and Characterization of MicroRNAs of Human Mature Erythrocytes 193
    14. Stable Overexpression of miRNAs in Bone Marrow-Derived Murine Mast Cells Using Lentiviral Expression Vectors
    15. Monitoring MicroRNA Activity and Validating MicroRNA Targets by Reporter-Based Approaches
    PART IV FUNCTIONAL ANALYSIS OF miRNAs IN THE IMMUNE SYSTEM: LOSS-OF-FUNCTION
    16. Lentivirus-Mediated Antagomir Expression
    PART V MIRNA POST-TRANSCRIPTIONAL MODIFICATIONS AND MFCHANISMS OF ACTION
    17. Solution Structure of miRNA: mRNA Complex
    18. MiRNA Editing
    PART VI BIOINFORMATIC ANALYSIS AND TARGET PREDICTION
    19. Computational Prediction of MicroRNA Targets
    20. Large-Scale Integration of MicroRNA and Gene Expression Data for Identification of Enriched MicroRNA-mRNA Associations in Biological Systems 297
    PART VII MIRNA AND VIRUSES
    21. Identification and Validation of the Cellular Targets of Virus-Encoded MicroRNAs
    Index 327