The Use Of Crispr Cas9 Zfns Talens In Generating Site Specific Genome Alterations

This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers recent research and methods development for changing the DNA sequence within the genomes of cells and organisms. Focusing on enzymes that generate double-strand breaks in DNA, the chapters describe use of molecular tools to introduce or delete genetic information at specific sites in the genomes of animal, plant and bacterial cells. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers research methods in biomineralization science Contains sections on such topics as genome editing, genome engineering, CRISPR, Cas9, TALEN and zinc finger nuclease

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.

CRISPR/Cas is a recently described defense system that protects bacteria and archaea against invasion by mobile genetic elements such as viruses and plasmids. A wide spectrum of distinct CRISPR/Cas systems has been identified in at least half of the available prokaryotic genomes. On-going structural and functional analyses have resulted in a far greater insight into the functions and possible applications of these systems, although many secrets remain to be discovered. In this book, experts summarize the state of the art in this exciting field.

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.

Recent advances in genome editing tools using endonucleases such as TALENs, ZFNs, and CRISPRs, combined with genomic engineering technologies, have opened up a wide range of opportunities from applications in the basic sciences and disease biology research, to the potential for clinical applications and the development of new diagnostic tools. This complete guide to endonuclease-based genomic engineering gives readers a thorough understanding of this rapidly expanding field. Chapters cover the discovery, basic science, and application of these techniques, focusing particularly on their potential relevance to the treatment of cancer, and cardiovascular and immunological disease. The final section discusses the legal and ethical issues which accompany the technology. Providing authoritative coverage of the potential that genome editing and engineering have, this is an ideal reference for researchers and graduate students and those working in the biotechnology and pharmaceutical industries, as well as in a clinical setting.

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.

Over the last few decades, various techniques have been developed to alter the properties of plants and animals. While the targeted transfer of recombinant DNA into crop plants remains a valuable tool to achieve a desirable breeding outcome, integration of transgenes into the host genome has been random, which in part, leads to reduced acceptance of GMOs by the general population in some parts of the world. Likewise, methods of induced mutagenesis, such as TILLING, have the disadvantage that many mutations are induced per plant, which has to be removed again by expensive backcrossing. Advances in genome sequencing have provided more and more information on differences between susceptible and resistant varieties, which can now be directly targeted and modified using CRISPR/Cas9 technology. By selecting specific gRNAs occurrence of off-target modifications are comparatively low. ZFNs and TALENs- based approaches required re-engineering a new set of assembled polypeptides for every new target site for each experiment. The difficulty in cloning and protein engineering prevented these tools from being broadly adopted by the scientific community. Compared to these technologies, designing the CRISPR toolbox is much simpler and more flexible. CRISPR/Cas9 is versatile, less expensive and highly efficient. It has become the most widely used technology for genome editing in many organisms. Since its inception as a powerful genome-editing tool in late 2012, this breakthrough technology has completely changed how science is performed. The first few chapters in this book introduce the basic concept, design and implementation of CRISPR/Cas9 for different plant systems. They are followed by in-depth discussions on the legal and bio-safety issues accompanying commercialization and patenting of this emerging technology. Lastly, this book covers emerging areas of new tools and potential applications. We believe readers, novice and expert alike, will benefit from this all-in-one resource on genome editing for crop improvement. Chapter 17 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This book discusses CRISPR/Cas- one of the most powerful tools available to scientists for genome editing. CRISPR/Cas is not only a genome editing tool, but researchers have also engineered it for gene regulation, genome imaging, base editing and epigenome regulations. This book describes the entire toolkit for CRISPR/Cas. The opening section gives an introduction to the technique and compares it with other genome editing tools. Further section gives a historical perspective of the tool, along with its detailed classification. The next chapters describe bioinformatic tools in CRISPR/Cas, and delivery methods for CRISPR/Cas. The book also discusses about the applications of CRISPR/Cas beyond genome editing and use of CRISPR for rewriting genetic codes. The book dedicates a section to the use of CRISPR in plants. The book culminates with a chapter on the current status, challenges and shortcomings of the CRISPR/Cas genome editing tool. The book would be highly interesting to students and researchers in molecular biology, biochemistry, biotechnology, food science, agriculture and plant sciences.

This volume details generation of gene-edited cell lines and organisms as models for human diseases, pest control, and large animal welfare and production outcomes. Chapters guide readers through gene regulation, editing, screening of cell lines, genome editing, and an overview of the tools for efficient genome editing including; ZFNs, TALENs, and CRISPR. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step- by-step, readily reproducible protocols. Authoritative and cutting-edge, Applications of Genome Modulation and Editing aims to be a useful and practical guide for researchers to commence or advance their study in this field.

Genetically engineered (GE) crops were first introduced commercially in the 1990s. After two decades of production, some groups and individuals remain critical of the technology based on their concerns about possible adverse effects on human health, the environment, and ethical considerations. At the same time, others are concerned that the technology is not reaching its potential to improve human health and the environment because of stringent regulations and reduced public funding to develop products offering more benefits to society. While the debate about these and other questions related to the genetic engineering techniques of the first 20 years goes on, emerging genetic-engineering technologies are adding new complexities to the conversation. Genetically Engineered Crops builds on previous related Academies reports published between 1987 and 2010 by undertaking a retrospective examination of the purported positive and adverse effects of GE crops and to anticipate what emerging genetic-engineering technologies hold for the future. This report indicates where there are uncertainties about the economic, agronomic, health, safety, or other impacts of GE crops and food, and makes recommendations to fill gaps in safety assessments, increase regulatory clarity, and improve innovations in and access to GE technology.