免疫学技术

Gene expression regulation by miRNAs has been reported to control key aspects of B cell differentiation and function (Chen et al., Science 303:83–86, 2004; Xiao et al., Cell 131:146–159, 2007; O’Carroll et al., Genes Dev. 21:1999–2004, 2007; Koralov et al. Cell 132:860–874, 2008; Rodriguez et al., Science ...

MicroRNAs (miRNAs or miRs) are ∼22 nt single-stranded noncoding RNAs that control gene expression in eukaryotes. miRNAs play an essential role in all basic cellular processes including cell development, proliferation, differentiation, and apoptosis. Importantly, miRNAs regu ...

In 1991, Soriano and coworkers isolated the ROSA26 locus in a gene-trap mutagenesis screening performed in mouse embryonic stem (ES) cells. The ubiquitous expression of ROSA26 in embryonic and adult tissues, together with the high frequency of gene-targeting events observed at this locus ...

The discovery of small regulatory RNA molecules during the last few years has changed our understanding of many biological and pathological processes. The most prominent and best analyzed class of these small regulatory noncoding RNAs is comprised by the microRNAs. The analysis of micro ...

Since the phenomenon of small RNA-mediated gene silencing was first described over 15 years ago (Lee et al. Cell 75:843–854, 1993; Wightman et al. Cell 75:855–862, 1993), it has become evident that a variety of endogenous small RNAs play an important role in establishing and maintaining cell lineage ...

The expression profile of microRNAs significantly varies in physiological and pathological conditions. Increasing evidence from the literature shows that abnormalities of the miRNome (defined as the full spectrum of miRNAs expressed in a genome) occur in almost all human diseases ...

MicroRNAs have emerged as – important posttranscriptional regulators of gene expression. Small RNA cloning is a powerful method to identify new microRNAs (miRNAs) and to profile miRNA expression. In addition, it reveals end heterogeneity that may be important in miRNA function. Here, we d ...

MicroRNAs (miRNAs) are small noncoding RNAs of an average length of 22 nucleotides, which repress translation of a large number of target mRNAs. The particular importance of this group of small RNAs arises from the ever growing evidence that they control many biological processes, such as diff ...

The protocol reported in this chapter describes a method for the detection and spatial localisation of microRNAs (miRNAs) in cryopreserved primary leukaemic suspension cells using digoxigenin (DIG)-labelled, Locked Nucleic Acid (LNA)-modified probes, and fluorescence in si ...

Proper normalization of quantitative RT-PCR (qRT-PCR) data is a crucial component of gene �expression analysis. Although arbitrarily selected housekeeping genes have been used to normalize many published mRNA RT-PCR datasets, there is a growing awareness that such normalizers sh ...

This protocol describes a method that uses splinted ligation for in-solution, direct labeling of small RNAs from total RNA. The liquid phase hybridization method makes it possible to achieve sensitive, specific, and quantitative detection while eliminating a number of time-consumi ...

Since the identification of the first virus-encoded microRNA (miRNA) in Epstein–Barr virus (EBV)-infected B cells in 2004, viral miRNAs have been found in different groups of herpesviruses. Viral miRNAs play an important role in regulating both viral and cellular gene expression. Ident ...

The discovery of microRNAs (miRNAs) revealed a hidden layer of gene regulation that is able to integrate multiple genes into biologically meaningful networks. A number of computational prediction programs have been developed to identify putative miRNA targets. Collectively, the m ...

One critical step in miRNA functional studies is to identify the gene targets that are directly regulated by miRNAs. In this chapter, we describe a computational algorithm and an online database, miRDB, for miRNA target prediction. In miRDB, flexible Web search interface has been developed for ...

RNA editing by A-to-I modification is a widespread mechanism in complex organisms that leads to the posttranscriptional alteration of protein coding as well as noncoding sequences. MiRNA transcripts have been recognized as a major target for RNA editing enzymes, and single-nucleoti ...

The use of contemporary nuclear magnetic resonance (NMR) methods in the studies of model systems between microRNA (miRNA) and messenger RNA (mRNA) is reviewed. We describe our studies on structural features of 33-nt RNA model construct between let-7 miRNA and lin-41 mRNA at the second binding s ...

MicroRNAs (miRNAs) are small non-coding RNAs involved in post-transcriptional gene regulation via hybridisation to mRNAs. miRNA function can be inhibited by the so-called “antagomirs” – anti-sense RNA oligonucleotides complementary to individual miRNAs. Since, in principle, ...

In this chapter, we provide a review on the functions of the most important miRNAs in lymphocytes. Most of them are involved in lymphopoiesis, immune response, and lymphoid malignancies, highlighting the importance of miRNAs in these cells.

An essential requirement for discovering microRNAs that may be relevant to an immune cell’s function is to identify the microRNAs that are active in the cell and the genes they target. As several chapters in this volume describe, there are a number of technologies available for profiling microR ...

Pathogen recognition is a central activity of the Toll-like receptor (TLR) family. Molecules from various pathogens have been widely used in TLR research as natural ligands for the receptors. TLR ligands from bacteria, viruses, and fungi are widely available from commercial companies and ...