Microrna Detection And Target Identification

This updated volume reflects new and evolved techniques to study detection, profiling, and manipulation of microRNAs (miRNAs) in plants and animals. After overviews of how best to detect, identify, and validate microRNAs, the book continues by exploring state-of-the-art protocols for microRNA detection, approaches to profile the expression level of microRNAs, spatial expression analysis, describe in silico analysis of microRNAs and their targets, as well as protocols for functional analysis of microRNAs and their targets by CRISPR/Cas. 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, MicroRNA Detection and Target Identification: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

MicroRNAs (miRNAs), endogenous noncoding regulatory mRNAs of - nucleotides, have rapidly emerged as the central players in gene expression regulation. Owing to their ever-increasing implications in the control of various biological and pathological processes, miRNAs have now been considered novel biomarkers of various human diseases including, cancer, viral disease, cardiov- cular disorders, metabolic disturbances, etc. Particular expression pro?les have been associated with particular pathological states. Expression pro?ling of miR- NAs have therefore become extremely important not only for fundamentalists but also for clinicians. However, the methodologies used for detecting protein-coding mRNAs cannot be directly applied to miRNAs because of their small size. Over the past years, researchers have made great efforts to developing techniques suitable for miRNA detection and quanti?cation; a wide spectrum of creative and inno- tive techniques (more than 30 different methods) have been invented and validated. It has come to the time now to summarize these methods and present them in an orderly manner for better understanding and utilization of these methods to miRNA research and applications. In particular, the development of methods for quantifying circulating miRNAs opens up a fascinating opportunity for realizing miRNA as diagnostic and prognostic biomarkers of human disease. A book on this subject may help boosting up the passion of researchers to further improve the existing techniques and develop more new methods to ?t to new application needs. These considerations prompted us and urged us to undertake the work: writing a book focusing on miRNA expression detection methods.

MicroRNAs (miRNA) are tiny bits of genetic material that were unknown nearly 10 years ago but now represent an exciting field of study in biology. Upon their discovery, researchers revealed for the first time a new mechanism by which microRNA can stop the function of messenger RNA (mRNA) by literally cutting it in half, interfering with the normal function of specific messenger RNAs in gene expression. This "expression" of genes that code for essential proteins is essentially what controls whether a cell turns into a liver, lung, or brain cell, for example. Understanding what activates this process – or stops it – is a key to understanding the biological process and builds a foundation for advances in medicine and other fields. This volume in Methods in Enzymology presents valuable methods for studying MicroRNA, with three sections covering identification of MicroRNAs and their targets; MicroRNA expression, maturation and functional analysis; and MicroRNAs and disease.

This book summarizes microRNA (miRNA) biology in a variety of pathological processes, emphasizing the significant potential applications of miRNA in diagnostics and prognostics, as well as novel drug targets. The conventional techniques used for miRNA detection including standard PCR, Northern blotting, microarray and clone methods are addressed. Recent emerging strategies in miRNA detection and quantification with superior flexibility and adaptability, such as novel molecular biological techniques and locked nucleic acid (LNA) modified probes, as well as nanotechnology-based approaches, are also included. The book also highlights the latest advances in clinical-related miRNA detection methods in living cells, circulating blood and tissue, such as in situ hybridization (ISH) and molecular imaging techniques, which are useful to elucidate the biogenesis and biological function of miRNAs in vivo. Finally, the respective advantages and drawbacks of various detection techniques in this fast-moving field are discussed, along with the challenges and promising new directions. This book offers a valuable resource for analytical chemists, biologists and physicians involved in miRNA research. Dr. Xueji Zhang and Dr. Haifeng Dong are Professors at the School of Chemistry & Biological Engineering, University of Science & Technology Beijing (USTB), China. Dr. Yaping Tian is a Professor at the Department of Clinical Biochemistry, Chinese PLA General Hospital and Military Medical School, China.

Extensive studies have been conducted on the identification, biogenesis, and processing of microRNA (miRNA) as well as research on the exact mechanism by which miRNAs bring about translational silencing of their targets. In addition, numerous publications point to an important role of miRNAs in development, reprogramming, epigenetics, pathogenesis of cancer, oncogenes and tumor suppressors, biomarkers of various disease onset, and regulation of adipogenesis and obesity; yet many questions still remain. In MicroRNA Protocols, Second Edition, experts in the field provide up-to-date coverage of this diverse area of study. The specific chapters of this edition are related to the analysis of miRNA, targets and expression profiling, various methods to determine its regulation of gene expression, the preparation and isolation of miRNAs in specific tissues, its detection in the saliva, and potential application in cosmetics, wound healing, and prostate cancer. Written in the highly successful Methods in Molecular BiologyTM series format, 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 known pitfalls. Fully updated and easy to use, MicroRNA Protocols, Second Edition aims to stimulate readers to explore diverse ways to understand the mechanisms in which miRNAs facilitate the molecular aspects of not only biomedical research but also a wide range of other research fields.

Since the discovery of microRNAs, developmental biologists have striven to understand the role of miRNAs in development and disease. MicroRNAs in Development: Methods and Protocols collects contributions from expert researchers in order to provide practical guidelines to this complex study. Divided into three convenient sections, this detailed volume covers various techniques to detect and profile miRNA expression, followed by protocols to manipulate the activity of miRNAs in various organisms, and it concludes with a section that outlines different methods to identify and validate miRNA targets in animals and plants. Written in the highly successful Methods in Molecular BiologyTM series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, MicroRNAs in Development: Methods and Protocols serves as a practical guide for scientists of all backgrounds and conveys the appropriate sense of fascination associated with this vital field of research.

MicroRNAs (miRNAs) are RNA molecules, conserved by evolution, that regulate gene expressions and their recent discovery is revolutionising both basic biomedical research and drug discovery. Expression levels of MiRNAs have been found to vary between tissues and with developmental stages and hence evaluation of the global expression of miRNAs potentially provides opportunities to identify regulatory points for many different biological processes. This wide-ranging reference work, written by leading experts from both academia and industry, will be an invaluable resource for all those wishing to use miRNA techniques in their own research, from graduate students, post-docs and researchers in academia to those working in R&D in biotechnology and pharmaceutical companies who need to understand this emerging technology. From the discovery of miRNAs and their functions to their detection and role in disease biology, this volume uniquely integrates the basic science with industry application towards drug validation, diagnostic and therapeutic development. Forewords by: Sidney Altman, Yale University, Winner of the Nobel Prize in Chemistry, 1989 and Victor R. Ambros, Dartmouth Medical School, Co-discoverer of MicroRNAs