Rna Tagging

This book provides a compendium of state-of-the-art methods for the labeling, detection, and purification of RNA and RNA-protein complexes and thereby constitutes an important toolbox for researchers interested in understanding the complex roles of RNA molecules in development, signaling, and disease. Beginning with a section on in situ detection of RNA molecules using FISH techniques, the volume continues with parts exploring in vivo imaging of RNA transport and localization, imaging and analysis of RNA uptake and transport between cells, identification and analysis of RNA-binding proteins, guide RNAs in genome editing, as well as other specific analytical techniques. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, RNA Tagging: Methods and Protocols serves as a vital reference for researchers looking to further the increasingly important research in RNA biology.

Specific complexes of protein and RNA carry out many essential biological functions, including RNA processing, RNA turnover, RNA folding, as well as the translation of genetic information from mRNA into protein sequences. Messenger RNA (mRNA) decay is now emerging as an important control point and a major contributor to gene expression. Continuing identification of the protein factors and cofactors, and mRNA instability elements responsible for mRNA decay allow researchers to build a comprehensive picture of the highly orchestrated processes involved in mRNA decay and its regulation. Covers the nonsense-mediated mRNA decay (NMD) or mRNA surveillance pathway Expert researchers introduce the most advanced technologies and techniques to identify mRNA processing, transport, localization and turnover, which are central to the process of gene expression Offers step-by-step lab instructions, including necessary equipment and reagents

The presence of modified nucleotides in cellular RNAs has been known for decades and over 100 distinct RNA modifications have been characterized to date. While the exact role of many of these modifications is still unclear, many are highly conserved across evolution and most contribute to the overall fitness of the organism. In recent years, new methods and bioinformatics approaches have been developed for the dissection of modification pathways and functions. These methods intersect a number of related fields, ranging from RNA processing to comparative genomics and systems biology. In addition, many of the techniques described in this volume have broad applicability, particularly in regards to the isolation, characterization, and reconstitution of ribonucleoprotein complexes, expanding the experimental repertoire available to all RNA researchers.

This MIE volume provides laboratory techniques that aim to predict the structure of a protein which can have tremendous implications ranging from drug design, to cellular pathways and their dynamics, to viral entry into cells. Expert researchers introduce the most advanced technologies and techniques in protein structure and folding Includes techniques on tiling assays

RNA and DNA Editing assembles a team of leading experts who present the latest discoveries in the field alongside the latest models and methodology. In addition, the authors set forth the many open questions and suggest routes for further investigation. Overall, the book serves as a practical guide for professionals in the field who need to understand the interrelationship of RNA and DNA editing with other chemical and biological processes.

This book presents an overview of the RNA networks controlling gene expression in fungi highlighting the remaining questions and future challenges in this area. It covers several aspects of the RNA-mediated mechanisms that regulate gene expression in model yeasts and filamentous fungi, organisms of great importance for industry, medicine and agriculture. It is estimated that there are more than one million fungal species on the Earth. Despite their diversity (saprophytic, parasitic and mutualistic), fungi share common features distinctive from plants and animals and have been grouped taxonomically as an independent eukaryotic kingdom. In this book, 15 chapters written by experts in their fields cover the RNA-dependent processes that take place in a fungal cell ranging from formation of coding and non-coding RNAs to mRNA translation, ribosomal RNA biogenesis, gene silencing, RNA editing and epigenetic regulation.

Transfer RNAs (tRNAs) are one of the classical non-coding RNAs whose lengths are approximately 70–100 bases. The secondary structure of tRNAs can be represented as the cloverleaf with 4 stems, and the three dimensional structure as an “L” shape. Historically, the basic function of tRNA as an essential component of translation was established in 1960s, i.e., each tRNA is charged with a target amino acid and these are delivered to the ribosome during protein synthesis. However, recent data suggests that the role of tRNA in cellular regulation goes beyond this paradigm. In most Archaea and Eukarya, precursor tRNAs are often interrupted by a short intron inserted strictly between the first and second nucleotide downstream of the anticodon, known as canonical nucleotide position (37/38). Recently, a number of reports describe novel aspects of tRNAs in terms of gene diversity, for example, several types of disrupted tRNA genes have been reported in the Archaea and primitive Eukarya, including multiple-intron-containing tRNA genes, split tRNA genes, and permuted tRNA genes. Our understanding of the enzymes involved in tRNA functions (e.g., aminoacyl-tRNA synthetase, tRNA splicing endonuclease, tRNA ligase) has deepened. Moreover, it is well known that tRNA possesses many types of base modifications whose enzymatic regulations remain to be fully elucidated. It was reported that impaired tRNA nuclear-cytoplasmic export links DNA damage and cell-cycle checkpoint. Furthermore, a variety of additional functions of tRNA, beyond its translation of the genetic code, have emerged rapidly. For instance, tRNA cleavage is a conserved part of the responses to a variety of stresses in eukaryotic cells. Age-associated or tissue-specific tRNA fragmentation has also been observed. Several papers suggested that some of these tRNA fragments might be involve in the cellular RNA interference (RNAi) system. These exciting data, have lead to this call for a Research Topic, that plans to revisit and summarize the molecular biology of tRNA. Beyond the topics outlined above, we have highlighted recent developments in bioinformatics tools and databases for tRNA analyses.