Nuclease Methods and Protocols
Nuclease Methods and Protocols

Author: Catherine H. Schein

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 521

ISBN-13: 1592592333

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Nucleases, enzymes that restructure or degrade nucleic acid polymers, are vital to the control of every area of metabolism. They range from “housekeeping” enzymes with broad substrate ranges to extremely specific tools (1). Many types of nucleases are used in lab protocols, and their commercial and clinical uses are expanding. The purpose of Nuclease Methods and Protocols is to introduce the reader to some we- characterized protein nucleases, and the methods used to determine their activity, structure, interaction with other molecules, and physiological role. Each chapter begins with a mini-review on a specific nuclease or a nuclease-related theme. Although many chapters cover several topics, they were arbitrarily divided into five parts: Part I, “Characterizing Nuclease Activity,” includes protocols and assays to determine general (processive, distributive) or specific mechanisms. Methods to assay nuclease products, identify cloned nucleases, and determine their physiological role are also included here. Part II, “Inhibitors and Activators of Nucleases,” summarizes assays for measuring the effects of other proteins and small molecules. Many of these inhibitors have clinical relevance. Part III, “Relating Nuclease Structure and Function,” provides an overview of methods to determine or model the 3-D structure of nucleases and their complexes with substrates and inhibitors. A 3-D structure can greatly aid the rational design of nucleases and inhibitors for specific purposes. Part IV, “Nucleases in the Clinic,” summarizes assays and protocols suitable for use with t- sues and for nuclease based therapeutics.

Molecular Biology of Nucleases
Molecular Biology of Nucleases

Author: Nawin C. Mishra

Publisher: CRC Press

Published: 1995-03-06

Total Pages: 300

ISBN-13: 9780849376580

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Nucleases occupy a central position in the biochemistry of DNA transactions and other metabolism of nucleic acids in all organisms. They have also proven useful in modern biological studies crucial for the development of recombinant DNA technology and reverse genetics. Nucleases assist in the identification and characterization of genes responsible for several diseases and their possible alleviation by gene therapy. Molecular Biology of Nucleases introduces the properties and biological roles of nucleases. It is the one comprehensive source for newcomers to the field.

Targeted Genome Editing Using Site-Specific Nucleases
Targeted Genome Editing Using Site-Specific Nucleases

Author: Takashi Yamamoto

Publisher: Springer

Published: 2015-01-05

Total Pages: 206

ISBN-13: 4431552278

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This book serves as an introduction to targeted genome editing, beginning with the background of this rapidly developing field and methods for generation of engineered nucleases. Applications of genome editing tools are then described in detail, in iPS cells and diverse organisms such as mice, rats, marine invertebrates, fish, frogs, and plants. Tools that are mentioned include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9, all of which have received much attention in recent years as breakthrough technologies. Genome editing with engineered nucleases allows us to precisely change the target genome of living cells and is a powerful way to control functional genes. It is feasible in almost all organisms ranging from bacteria to plants and animals, as well as in cultured cells such as ES and iPS cells. Various genome modifications have proven successful, including gene knockout and knock-in experiments with targeting vectors and chromosomal editing. Genome editing technologies hold great promise for the future, for example in biomedical research, clinical medicine, and generation of crops and livestock with desirable traits. A wide range of readers will find this book interesting, and with its focus on applications in a variety of organisms and cells, the book will be valuable for life scientists in all fields.

Artificial Nucleases
Artificial Nucleases

Author: Marina A. Zenkova

Publisher: Springer Science & Business Media

Published: 2004-01-07

Total Pages: 330

ISBN-13: 9783540201120

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The development of agents capable of cleaving RNA and DNA has attracted considerable attention from researchers in the last few years, because of the immediate and very important applications they can find in the emerging fields of biotechnology and pharmacology. There are essentially two classes of these agents - nucleases that occur naturally inside cells and synthetically produced artificial nucleases. The first class includes protein enzyme nucle ases and catalytic RNA structured ribozymes that perform cleavage of the phosphodiester bonds in nucleic acids according to a hydrolytic pathway in the course of different biochemical processes in the cell. A different pathway is used by some antibiotics which cleave DNA via redox-based mechanisms resulting in oxidative damage of nucleotide units and breakage of the DNA backbone. The above molecules are indispensable tools for manipulating nucleic acids and processing RNA; DNA-cleaving antibiotics and cytotoxic ribonucleases have demonstrated utility as chemotherapeutic agents. The second class, artificial nucleases, are rationally designed to imitate the active centers of natural enzymes by simple structures possessing minimal sets of the most important characteristics that are essential for catalysis. A dif ferent approach, in vitro selection, was also used to create artificial RNA and DNA enzymes capable of cleaving RNA. Being less efficient and specific as compared to the natural enzymes, the primitive mimics are smaller and robust and can function in a broad range of conditions.

Comparative Virology
Comparative Virology

Author: Karl Maramorosch

Publisher: Academic Press

Published: 2014-06-28

Total Pages: 601

ISBN-13: 1483269698

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Comparative Virology provides an integrated comparison of viruses, based on their chemical and morphological characteristics. These descriptions will not only give the reader a background but also a detailed analysis of the various groups. In some instances the groups are still host related, as in the case of bacteriophages and polyhedral insect viruses. In others, for instance in pox viruses, the group comprises viruses of vertebrates and invertebrates. The hosts of the bacilliform Rhabdovirales range from man and other warm-blooded vertebrates through invertebrate animals to plants. A special chapter is devoted to viruses devoid of protein—a group that is of great interest and that has only recently been recognized. Since there is historical and practical interest in écologie groupings, such as arboviruses and oncogenic viruses, chapters on such groups have also been included. The book opens with a discussion on the classification of viruses. Chapters dealing with DNA viruses and RNA viruses follow, and the ecologically and disease-oriented groups complete the volume. It is hoped that ""Comparative Virology"" will help bring unity to the science of virology through the comparative approach that is not dependent on virus-host interactions. The combined efforts of eminent contributors to discuss and evaluate new information will hopefully benefit all who are interested in virology

Restriction Endonucleases
Restriction Endonucleases

Author: Alfred Pingoud

Publisher: Springer Science & Business Media

Published: 2012-12-06

Total Pages: 462

ISBN-13: 3642188516

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Restriction enzymes are highly specific nucleases which occur ubiquitously among prokaryotic organisms, where they serve to protect bacterial cells against foreign DNA. Many different types of restriction enzymes are known, among them multi-subunit enzymes which depend on ATP or GTP hydrolysis for target site location. The best known representatives, the orthodox type II restriction endonucleases, are homodimers which recognize palindromic sequences, 4 to 8 base pairs in length, and cleave the DNA within or immediately adjacent to the recognition site. In addition to their important biological role (up to 10 % of the genomes of prokaryotic organisms code for restriction/modification systems!), they are among the most important enzymes used for the analysis and recombination of DNA. In addition, they are model systems for the study of protein-nucleic acids interactions and, because of their ubiquitous occurence, also for the understanding of the mechanisms of evolution.

RNA Turnover in Eukaryotes: Nucleases, Pathways and Analysis of mRNA Decay
RNA Turnover in Eukaryotes: Nucleases, Pathways and Analysis of mRNA Decay

Author: Lynne E. Maquat

Publisher: Academic Press

Published: 2009-01-30

Total Pages: 661

ISBN-13: 0080922074

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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

Gene Editing in Plants
Gene Editing in Plants

Author:

Publisher: Academic Press

Published: 2017-07-14

Total Pages: 266

ISBN-13: 0128117443

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Gene Editing in Plants, Volume 149 aims to provide the reader with an up-to-date survey of cutting-edge research with gene editing tools and an overview of the implications of this research on the nutritional quality of fruits, vegetables and grains. New chapters in the updated volume include topics relating to Genome Engineering and Agriculture: Opportunities and Challenges, the Use of CRISPR/Cas9 for Crop Improvement in Maize and Soybean, the Use of Zinc-Finger Nucleases for Crop Improvement, Gene Editing in Polyploid Crops: Wheat, Camelina, Canola, Potato, Cotton, Peanut, Sugar Cane, and Citrus, and Gene Editing With TALEN and CRISPR/Cas in Rice. This ongoing serial contain contributions from leading scientists and researchers in the field of gene editing in plants who describe the results of their own research in this rapidly expanding area of science. - Shows the importance of revolutionary gene editing technology on plant biology research and its application to agricultural production - Provides insight into what may lie ahead in this rapidly expanding area of plant research and development - Contains contributions from major leaders in the field of plant gene editing

Human Genome Editing
Human Genome Editing

Author: National Academies of Sciences, Engineering, and Medicine

Publisher: National Academies Press

Published: 2017-08-13

Total Pages: 329

ISBN-13: 0309452880

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Genome editing is a powerful new tool for making precise alterations to an organism's genetic material. Recent scientific advances have made genome editing more efficient, precise, and flexible than ever before. These advances have spurred an explosion of interest from around the globe in the possible ways in which genome editing can improve human health. The speed at which these technologies are being developed and applied has led many policymakers and stakeholders to express concern about whether appropriate systems are in place to govern these technologies and how and when the public should be engaged in these decisions. Human Genome Editing considers important questions about the human application of genome editing including: balancing potential benefits with unintended risks, governing the use of genome editing, incorporating societal values into clinical applications and policy decisions, and respecting the inevitable differences across nations and cultures that will shape how and whether to use these new technologies. This report proposes criteria for heritable germline editing, provides conclusions on the crucial need for public education and engagement, and presents 7 general principles for the governance of human genome editing.