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The recent expansion in diversity of RNA and DNA editing types has stimulated the development of many unique genetic, molecular, biochemical, and computational approaches to biological issues. In RNA and DNA Editing: Methods and Protocols, leading experts in the field introduce methods developed over the last few years to study editing substrates, mechanisms of specificity, and functions of RNA and DNA editing enzymes and complexes. Sections of the book are dedicated to state-of-the art techniques which enable investigation of uracil insertion/deletion RNA editing in mitochondrion of Trypanosoma brucei, adenosine to inosine RNA editing, cytidine to uracil RNA and DNA editing, as well as tRNA editing and RNA modifications. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, RNA and DNA Editing: Methods and Protocols seeks to inspire the further development of these vital and powerful techniques.
This volume presents a comprehensive collection of cuttingedge methods for elucidating the function of new genes and altering gene expression. These readily reproducible techniques can be used either in transient and stable gene splicing applied to worms, flies, trypanosomes, mammals, and plants, or in studying RNA editing mechanisms in a wide range of organisms, including systems that involve the conversion of one base to another and insertion/deletion editing. Topics of interest include stable and transient RNA interference, gene silencing, RNA editing, bioinformatics, small noncoding RNAs, and RNomics. Special attention is given to methods for the identification and characterization of small RNAs involved in RNA interference or modification. Readily reproducible protocols for discovering new genes or altering gene expression.
RNA Modification Enzymes, Volume 659 in the Methods in Enzymology series, highlights new advances in the field, with this new volume presenting interesting chapters on a variety of related topics, including Locating chemical modifications in RNA sequences through ribonucleases and LC-MS based analysis, Development of RNA modification mapping pipelines for high-throughput sequencing approaches, AlkAniline-Seq for high-resolution mapping RNA m7G and m3C modifications, Facile detection of RNA phospho-methylation in cells, Detection and analysis of glycosylated queuosine modifications, A comprehensive pipeline for analysis of RNA 3’-end modification, Analysis of the epitranscriptome with ion-pairing reagent free oligonucleotide mass spectrometry, and more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology series Updated release includes the latest information on the RNA Modification Enzymes
Heritable human genome editing - making changes to the genetic material of eggs, sperm, or any cells that lead to their development, including the cells of early embryos, and establishing a pregnancy - raises not only scientific and medical considerations but also a host of ethical, moral, and societal issues. Human embryos whose genomes have been edited should not be used to create a pregnancy until it is established that precise genomic changes can be made reliably and without introducing undesired changes - criteria that have not yet been met, says Heritable Human Genome Editing. From an international commission of the U.S. National Academy of Medicine, U.S. National Academy of Sciences, and the U.K.'s Royal Society, the report considers potential benefits, harms, and uncertainties associated with genome editing technologies and defines a translational pathway from rigorous preclinical research to initial clinical uses, should a country decide to permit such uses. The report specifies stringent preclinical and clinical requirements for establishing safety and efficacy, and for undertaking long-term monitoring of outcomes. Extensive national and international dialogue is needed before any country decides whether to permit clinical use of this technology, according to the report, which identifies essential elements of national and international scientific governance and oversight.
RNA Modification provides a useful examination of the science and its role in biological regulation, the current frontier of life science research, and includes various RNA modications and their role in gene expression. It represents the most up-to-date knowledge and protocols available today. Dynamic RNA modifications and their roles in biological regulation are the current frontier of life science research This volume of Methods in Enzymology represents up to date knowledge and protocols
This book is open access under a CC BY 4.0 license. CRISPR-Cas9 is a rapid, efficient, versatile and relatively cheap method for dissecting the molecular pathways that are the basis of life, as well as for investigating and potentially rectifying faults in these pathways that result in disease. This book reviews how CRISPR-Cas9 and other genome editing techniques are advancing our understanding of development and function in the nervous system, uncovering the molecular causes of neurological disorders and providing tools for gene therapy.
Concerted efforts to deepen understanding of RNA modifications and their role in living systems hold the potential to advance human health, improve crop yields, and address other pressing societal challenges. RNA, which carries the information encoded by DNA to the places where it is needed, is amazingly diverse and dynamic. RNA is processed and modified through natural biological pathways, giving rise to hundreds, in some cases thousands, of distinct RNA molecules for each gene, thereby diversifying genetic information. RNA modifications are known to be pivotal players in nearly all biological processes, and their dysregulation has been implicated in a wide range of human diseases and disorders. Yet, our knowledge of RNA modifications remains incomplete, hindered by current technological limitations. Existing methods cannot discover all RNA modifications, let alone comprehensively sequence them on every RNA molecule. Nonetheless, what is known about RNA modifications has already been leveraged in the development of vaccines that helped saved millions of lives worldwide during the COVID-19 pandemic. RNA modifications also have applications beyond health, for example, enhancing agricultural productivity. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine calls for a focused, large-scale effort to accelerate technological innovation to harness the full potential of RNA modifications to address pressing societal challenges in health, agriculture, and beyond. This report assesses the scientific and technological breakthroughs, workforce, and infrastructure needs to sequence RNA and its modifications, and ultimately understand the roles RNA modifications play in biological processes and disease. It proposes a roadmap of innovation that will make it possible for any RNA from any biological system to be sequenced end-to-end with all of its modifications - a capability that could lead to more personalized and targeted treatments and instigate transformative changes across various sectors beyond health and medicine.