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| crispr cas9 guide rna design: CRISPR Guide RNA Design Tudor A. Fulga, David J. H. F. Knapp, Quentin R. V. Ferry, 2020-11-19 This detailed volume focuses on the CRISPR-associated guide RNA and how it can be designed, modified, and validated for a broad repertoire of purposes. Beginning with a section on computational design of target-specific guide RNAs, the book continues by covering chemical modifications to alter guide RNA stability, specificity, and efficiency, as well as to create inducible guide RNAs, append additional functional domains, and express guide RNAs in a conditional manner. It concludes with methods for measuring off-target guide RNA activity. 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 protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and essential, CRISPR Guide RNA Design: Methods and Protocols provides a comprehensive pipeline for guide RNA design and aims to be an invaluable resource in applying this powerful technology to basic research and therapeutic applications. |
| crispr cas9 guide rna design: CRISPR-Cas Systems Rodolphe Barrangou, John van der Oost, 2012-12-13 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. |
| crispr cas9 guide rna design: Genome Engineering via CRISPR-Cas9 System Vijai Singh, Pawan K. Dhar, 2020-02-21 Genome Engineering via CRISPR-Cas9 Systems presents a compilation of chapters from eminent scientists from across the globe who have established expertise in working with CRISPR-Cas9 systems. Currently, targeted genome engineering is a key technology for basic science, biomedical and industrial applications due to the relative simplicity to which they can be designed, used and applied. However, it is not easy to find relevant information gathered in a single source. The book contains a wide range of applications of CRISPR in research of bacteria, virus, algae, plant and mammalian and also discusses the modeling of drosophila, zebra fish and protozoan, among others. Other topics covered include diagnosis, sensor and therapeutic applications, as well as ethical and regulatory issues. This book is a valuable source not only for beginners in genome engineering, but also researchers, clinicians, stakeholders, policy makers, and practitioners interested in the potential of CRISPR-Cas9 in several fields. |
| crispr cas9 guide rna design: CRISPR Gene Editing Yonglun Luo, 2019-04-23 This detailed volume guides readers through strategic planning and user-friendly guidelines in order to select the most suitable CRISPR-Cas system and target sites with high activity and specificity. Methods covering CRISPR gRNA design, CRISPR delivery, CRISPR activity quantification (indel quantification), and examples of applying CRISPR gene editing in human pluripotent stem cells, primary cells, gene therapy, and genetic screening are included. 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 invaluable, CRISPR Gene Editing: Methods and Protocols will assist undergraduates, graduates, and researchers with detailed guidelines and methods for the vitally important CRISPR gene editing field. Chapter 3 is available open access under a CC BY 4.0 license via link.springer.com. |
| crispr cas9 guide rna design: RNA Nanotechnology Bin Wang, 2014-04-02 In the past few decades there has been incredible growth in bionano-related research, which has been accompanied by numerous publications in this field. Although various compilations address topics related to deoxyribonucleic acid (DNA) and protein, there are few books that focus on determining the structure of ribonucleic acid (RNA) and using RNA as building blocks to construct nanoarchitectures for biomedical and healthcare applications. RNA Nanotechnology is a comprehensive volume that details both the traditional approaches and the latest developments in the field of RNA-related technology. This book targets a wide audience: a broad introduction provides a solid academic background for students, researchers, and scientists who are unfamiliar with the subject, while the in-depth descriptions and discussions are useful for advanced professionals. The book opens with reviews on the basic aspects of RNA biology, computational approaches for predicting RNA structures, and traditional and emerging experimental approaches for probing RNA structures. This section is followed by explorations of the latest research and discoveries in RNA nanotechnology, including the design and construction of RNA-based nanostructures. The final segment of the book includes descriptions and discussions of the potential biological and therapeutic applications of small RNA molecules, such as small/short interfering RNAs (siRNAs), microRNAs (miRNAs), RNA aptamers, and ribozymes. |
| crispr cas9 guide rna design: CRISPR-Cas Jennifer A. Doudna, Prashant Mali, 2016 CRISPR/Cas-based techniques are revolutionizing the way geneticists and molecular biologists modify DNA sequences and modulate gene expression in cells and organisms. This laboratory manual presents step-by-step protocols for applying this cutting-edge technology to any system of interest. Contributors describe approaches for de. |
| crispr cas9 guide rna design: Oligonucleotide Synthesis Piet Herdewijn, 2008-02-04 A collection of powerful new techniques for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are a novel two-step process that yields a high purity, less costly, DNA, the synthesis of phosphorothioates using new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, and triple helix formation. The applications include using molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules of DNA, nucleic acid libraries, and the use of small interference RNA (siRNA) as an inhibitor of gene expression. |
| crispr cas9 guide rna design: Human Genome Editing National Academies of Sciences, Engineering, and Medicine, National Academy of Medicine, National Academy of Sciences, Committee on Human Gene Editing: Scientific, Medical, and Ethical Considerations, 2017-08-13 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. |
| crispr cas9 guide rna design: CRISPR-Cas Enzymes , 2019-01-25 CRISPR-Cas Enzymes, Volume 616, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Topics covered in this release include CRISPR bioinformatics, A method for one-step assembly of Class 2 CRISPR arrays, Biochemical reconstitution and structural analysis of ribonucleoprotein complexes in Type I-E CRISPR-Cas systems, Mechanistic dissection of the CRISPR interference pathway in Type I-E CRISPR-Cas system, Site-specific fluorescent labeling of individual proteins within CRISPR complexes, Fluorescence-based methods for measuring target interference by CRISPR-Cas systems, Native State Structural Characterization of CRISRP Associated Complexes using 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 CRISPR-Cas Enzymes |
| crispr cas9 guide rna design: RNA Tagging Manfred Heinlein, 2021-08-08 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. |
| crispr cas9 guide rna design: DNA-Protein Interactions Andrew Arthur Travers, 1993-04-30 The binding of proteins to DNA and the manipulation of DNA by proteins are crucial aspects of the biological role of DNA in the living cell. This book provides a comprehensive and lucid discussion of the molecular interactions involved. |
| crispr cas9 guide rna design: Gene Editing in Plants , 2017-07-14 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 |
| crispr cas9 guide rna design: Plant Genome Editing with CRISPR Systems Yiping Qi, 2019-01-05 This volume provides readers with wide-ranging coverage of CRISPR systems and their applications in various plant species. The chapters in this book discuss topics such as plant DNA repair and genome editing; analysis of CRISPR-induced mutations; multiplexed CRISPR/Cas9 systems; CRISPR-Cas12a (Cpf1) editing systems; and non-agrobacterium based CRISPR delivery systems. Written in the highly successful Methods in Molecular Biology 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 avoiding known pitfalls. Comprehensive and thorough, Plant Genome Editing with CRISPR Systems: Methods and Protocols is a valuable resource for any researcher interested in learning about and using CRISPR systems in plants. |
| crispr cas9 guide rna design: Yarrowia Lipolytica Ian Wheeldon, Mark Blenner, 2021 This volume provides a collection of protocols for the most common experimental methods used for engineering Yarrowia lipolytica. Chapters detail the basic theories underlying the methods described in each chapter. Written in the highly successful Methods in Molecular Biology 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 avoiding known pitfalls. Authoritative and cutting-edge, Yarrowia lipolytica: Methods and Protocols aims to provide experimentalists with a full account of the practical steps necessary for carrying out each protocol successfully. |
| crispr cas9 guide rna design: Computational Methods in Synthetic Biology Mario Andrea Marchisio, 2021-11-27 This second edition book provides complete coverage of the computational approaches currently used in Synthetic Biology. New chapters detail computational methods and algorithms for the design of bio-components, insight on CAD programs, analysis techniques, and distributed systems. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Computational Methods in Synthetic Biology, Second Edition aims to feature a broad overview of the research areas that can be met in the area of in silico Synthetic Biology. |
| crispr cas9 guide rna design: Rice Functional Genomics Narayana M. Upadhyaya, 2010-10-29 Rice has been chosen as a model cereal for functional genomics because of its well understood genetics. In addition, there is great similarities among all the cereals and grasses: any understanding of rice genes is directly applicable to the genes of other cereals. This book details rice functional genomics. It takes a multi-pronged genome-wide approach using structural similarities, expression profiles, and mutant phenotypes. Coverage includes the current status of genome sequencing and annotation, various tools, and resources being developed worldwide. |
| crispr cas9 guide rna design: Genome Editing and Engineering Krishnarao Appasani, 2018-08-23 A complete guide to endonuclease-based genomic engineering, from basic science to application in disease biology and clinical treatment. |
| crispr cas9 guide rna design: CRISPR in Animals and Animal Models , 2017-11-10 CRISPR in Animals and Animal Models, Volume 152, the latest release in the Progress in Molecular Biology and Translational Science series, explores the genome editing CRISPR system in cells and animal models, its applications, the uses of the CRISPR system, and the past, present and future of CRISPR genome editing. Topics of interest in this updated volume include a section on CRISPR history, The genome editing revolution, Programming CRISPR and its applications, CRISPR Delivery methods, CRISPR libraries and screening, CRISPR investigation in haploid cells, CRISPR in the generation of transgenic animals, CRISPR therapeutics, and Promising strategies and present challenges. Accessible to students and researchers alike Written by leading authorities in the field |
| crispr cas9 guide rna design: Synthetic Genomics Miguel Fernández-Niño, Luis H. Reyes, 2022-02-02 The current advances in sequencing, data mining, DNA synthesis, cloning, in silico modeling, and genome editing have opened a new field of research known as Synthetic Genomics. The main goal of this emerging area is to engineer entire synthetic genomes from scratch using pre-designed building blocks obtained by chemical synthesis and rational design. This has opened the possibility to further improve our understanding of genome fundamentals by considering the effect of the whole biological system on biological function. Moreover, the construction of non-natural biological systems has allowed us to explore novel biological functions so far not discovered in nature. This book summarizes the current state of Synthetic Genomics, providing relevant examples in this emerging field. |
| crispr cas9 guide rna design: Reprogramming the Genome: Applications of CRISPR-Cas in non-mammalian systems part A , 2021-03-28 Reprogramming the Genome: Applications of CRISPR-Cas in Non-mammalian Systems, Part A presents a collation of chapters written by global, eminent scientists. CRISPR-Cas9 system is an RNA-mediated immune system of bacteria and archaea that protects from bacteriophage infections. It is one of the revolutionized technologies to uplift biology to the next stages. Chapters in this release include An Introduction and applications of CRISPR-Cas Systems, History, evolution and classification of CRISPR-Cas associated systems, CRISPR based bacterial genome editing and removal of pathogens, CRISPR based genome editing and removal of human viruses, CRISPR based development of RNA editing and diagnostic platform, and much more. Additional sections cover Genome engineering in insects for control of vector borne diseases, Development of insect cell line using CRISPR technology, CRISPRing protozoan parasites to better understand the biology of diseases, CRISPR based genome editing of Caenorhabditis elegans, and a variety of other important topics. - Offers a basic understanding and clear picture of genome editing CRISPR-Cas systems in different organisms - Explains how to create an animal model for disease diagnosis/research and reprogram CRISPR for removal of virus, bacteria, fungi, protozoan, and many more - Discusses the advances, patents, applications, challenges and opportunities in CRISPR-Cas9 systems in basic sciences, biomedicine, virology, bacteriology, molecular biology, and many more |
| crispr cas9 guide rna design: CRISPR Magnus Lundgren, Emmanuelle Charpentier, Peter C. Fineran, 2015-05-20 This volume presents a list of cutting-edge protocols for the study of CRISPR-Cas defense systems and their applications at the genomic, genetic, biochemical and structural levels. CRISPR: Methods and Protocols guides readers through techniques that have been developed specifically for the analysis of CRISPR-Cas and techniques adapted from standard protocols of DNA, RNA and protein biology. Written in the highly successful Methods in Molecular Biology 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 avoiding known pitfalls. Authoritative and cutting-edge, CRISPR: Methods and Protocols provides a broad list of tools and techniques to study the interdisciplinary aspects of the prokaryotic CRISPR-Cas defense systems. |
| crispr cas9 guide rna design: The CRISPR/Cas System Muhammad Jamal, 2017 The use of CRISPR/Cas technology for genome editing suggests many potential applications, including the alteration of the germline of humans, animals and food crops. The speed and efficiency of the CRISPR/Cas system make it a potentially useful system for gene therapy. In this volume expert international authors provide a useful and timely review of the applications of the CRISPR/Cas system across diverse fields and explore further avenues and research directions of this novel and powerful editing technology. The technology and its application are reviewed with respect to reproduction and development, immunity and genetic diseases, system structure and system specificity. Some of the potential problems of the CRISPR/Cas system are also discussed, in particular the specificity of the system: this remains an important topic as improvement could lead to the more direct and efficient use of the CRISPR/Cas system in clinical settings. The authors also debate ethical concerns associated with this powerful new technology. This volume is a rigorous review of the applications and new opportunities for the CRISPR/Cas system and provides a stimulus for current and future research. An invaluable guide for all scientists working in the fields of genome editing and gene therapy the book is also recommended for all life sciences libraries. |
| crispr cas9 guide rna design: Genome Engineering for Crop Improvement Bidyut Kumar Sarmah, Basanta Kumar Borah, 2021-04-18 This book serves the teachers, researchers and the students as a handy and concise reference as well as guidebook while designing and planning for use of the advanced technologies for crop improvement. The content of the book is designed to cover the latest genome engineering techniques for crop improvement. The conventional breeding has got its limitations such as non-availability of desired genes within the genepool. In many cases, breeding has been highly used and it has nearly reached its highest limit so far as the productivity and production of crops are concerned. However, with increasing need of food and decreasing resources, including water, land, labour, etc., to feed the growing population, the alternative available ways of increasing crop productivity need to be explored and exploited. Genome engineering has a wide scope that includes technologies such as genetic engineering and transgenesis, RNA technologies, CRISPR, cisgenics and subgenics for better productivity and more efficient biotic and abiotic stress management. Therefore, the book is planned to enlighten the readers with the advanced technologies with examples and case studies, whenever possible. Efforts will be made to emphasize on general efforts on various major food crops; however, it would also be made clear that such efforts could be taken as proofs of concepts and that this could be extrapolated keeping the demand in mind. |
| crispr cas9 guide rna design: CRISPR Rodolphe Barrangou, Erik J. Sontheimer, Luciano A. Marraffini, 2022-09-14 “An excellent compendium of all things CRISPR from some of the leading minds in the field. With thorough coverage from every angle and beautifully detailed illustrations, this book is not to be missed!” Jennifer A. Doudna, Professor of Chemistry, Biochemistry & Molecular Biology, UC Berkeley; Founder, Innovative Genomics Institute; Nobel Laureate and coinventor of CRISPR technology “This journey through CRISPR biology and several of its breakthrough applications offers an exciting glimpse into one of the most beautiful and compelling fields in the life sciences.” David R. Liu, Director of the Merkin Institute at the Broad Institute of MIT and Harvard; Professor of Chemistry and Chemical Biology at Harvard University; coinventor of base editing and prime editing “A must read! The CRISPR topics, written by world-leading experts, span from the fascinating mechanistic underpinnings to the ingenious applications. One can read from start to finish or pick and choose themes. Either way, the book delivers utterly enjoyable learning!” Bonnie Bassler, Squibb Professor and Chair, Princeton University Department of Molecular Biology; Howard Hughes Medical Institute Investigator CRISPR-Cas systems have revolutionized the science of gene editing and their possible applications continue to expand, from basic research to potentially groundbreaking medical and commercial uses. Led by a distinguished team of editors, CRISPR: Biology and Applications explores the subject matter needed to delve into this fascinating area. Topics covered include: Classification and molecular mechanisms of CRISPR-Cas systems CRISPR-Cas evolution, regulation, expression, and function Uses for gene editing and modulation of gene expression CRISPR-based antimicrobials and phage resistance for medical and industrial purposes Written by internationally renowned authors, CRISPR: Biology and Applications serves as both an introductory guide for those new to the field and an authoritative reference for seasoned researchers whose work touches this evolving and headline- making science. |
| crispr cas9 guide rna design: Mammalian Cell Engineering Ryosuke Kojima, 2022-07-21 This volume explores the latest engineering methods of mammalian cells that are useful for controlling the performance of engineered mammalian cells for future cell-based therapeutics and for better understanding of complex biological systems. The chapters in this book are organized into five parts. Part One described methods to engineer mammalian cells to sense biologically relevant inputs, such as cell contacts and soluble proteins. Part Two looks at techniques to engineer mammalian cells to sense artificial inputs, such as light and ultrasound. Part Three provides cutting-edge CRISPR-Cas-based methods to carry out highly multiplexed genome editing and spatiotemporally controlled genome editing. Part Four discusses ways to control and engineer biological events in mammalian cells in combination with chemical compounds and systems. Part Five explores techniques to engineer specific mammalian cells in targeted manners. Written in the highly successful Methods in Molecular Biology 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 avoiding known pitfalls. Comprehensive and authoritative, Mammalian Cell Engineering: Methods and Protocols is a valuable resource that allows scientists to successfully carry out their research, thus ultimately contributing to the future advancement of this field. |
| crispr cas9 guide rna design: Microinjection David J. Carroll, 2008-12-12 As the number of sequenced genomes continues to increase, understanding the functions of newly discovered molecules will require greater efficiency and further study within the context of live cells. In Microinjection: Methods and Applications, expert researchers contribute methods utilizing microinjection techniques ranging from expression of RNA to the integration of DNA into the genome with the ultimate goal of learning about gene expression, signal transduction, and protein function within these living cells. This versatile volume updates established techniques such as cRNA expression in Xenopus oocytes, and examines new, cutting-edge technologies, including antisense morpholino oligonucleotides, RNAi for knockdown experiments, and the use of integrase to produce transgenic animals, all through microinjection techniques that can be easily adopted by any lab. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections, highlighting tips on troubleshooting and avoiding known pitfalls. Comprehensive and easily accessible, Microinjection: Methods and Applications is an ideal source for new ideas and techniques certain to aid in the advance of biological research. |
| crispr cas9 guide rna design: Bioinformatics in Agriculture Pradeep Sharma, Dinesh Yadav, R.K. Gaur, 2022-04-28 Bioinformatics in Agriculture: Next Generation Sequencing Era is a comprehensive volume presenting an integrated research and development approach to the practical application of genomics to improve agricultural crops. Exploring both the theoretical and applied aspects of computational biology, and focusing on the innovation processes, the book highlights the increased productivity of a translational approach. Presented in four sections and including insights from experts from around the world, the book includes: Section I: Bioinformatics and Next Generation Sequencing Technologies; Section II: Omics Application; Section III: Data mining and Markers Discovery; Section IV: Artificial Intelligence and Agribots. Bioinformatics in Agriculture: Next Generation Sequencing Era explores deep sequencing, NGS, genomic, transcriptome analysis and multiplexing, highlighting practices forreducing time, cost, and effort for the analysis of gene as they are pooled, and sequenced. Readers will gain real-world information on computational biology, genomics, applied data mining, machine learning, and artificial intelligence. This book serves as a complete package for advanced undergraduate students, researchers, and scientists with an interest in bioinformatics. - Discusses integral aspects of molecular biology and pivotal tool sfor molecular breeding - Enables breeders to design cost-effective and efficient breeding strategies - Provides examples ofinnovative genome-wide marker (SSR, SNP) discovery - Explores both the theoretical and practical aspects of computational biology with focus on innovation processes - Covers recent trends of bioinformatics and different tools and techniques |
| crispr cas9 guide rna design: Embryo Experimentation Peter Singer, Stephen Buckle, 1993 New developments in reproductive technology have made headlines since the birth of the world's first in vitro fertilization baby in 1978. But is embryo experimentation ethically acceptable? What is the moral status of the early human embryo? And how should a democratic society deal with so controversial an issue, where conflicting views are based on differing religious and philosophical positions? These controversial questions are the subject of this book, which, as a current compendium of ideas and arguments on the subject, makes an original contribution of major importance to this debate. Peter Singer is the author of many books, including Practical Ethics (CUP, 1979), Marx (Hill & Wang, 1980), and Should the Baby Live? (co-authored with Helga Kuhse, Oxford U.P., 1986). |
| crispr cas9 guide rna design: Heritable Human Genome Editing The Royal Society, National Academy of Sciences, National Academy of Medicine, International Commission on the Clinical Use of Human Germline Genome Editing, 2021-01-16 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. |
| crispr cas9 guide rna design: Modern Prometheus Jim Kozubek, 2016-10-18 This book tells the dramatic story of Crispr and the potential impact of this gene-editing technology. |
| crispr cas9 guide rna design: Plant Amino Acids Bijay K. Singh, 1998-10-27 Covers the basic knowledge of the regulation of biosynthesis of various amino acids in plants and the application of this knowledge to the discovery of novel inhibitors of amino acid biosynthesis and for enhancing the nutritional value of plant products. Provides an exhaustive list of pathway inhibitors. |
| crispr cas9 guide rna design: Pichia Protocols James M Cregg, 2007-08-08 This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition. |
| crispr cas9 guide rna design: Rice Grain Quality Nese Sreenivasulu, 2019-12-10 This volume explores the latest advances in high-throughput phenotyping to screen milling quality, cooking and nutritional quality in breeding material, and simplifying grain quality screening in germplasm. The chapters in this book cover topics such as seed development stages; panicle architectural traits to understand yield components; starch structure and protein properties; and genome editing technologies to reveal grain quality and nutritional traits. Written in the highly successful Methods in Molecular Biology 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 avoiding known pitfalls. Cutting-edge and thorough, Rice Grain Quality: Methods and Protocols is a valuable resource for all researchers interested in expanding their knowledge in this evolving field. |
| crispr cas9 guide rna design: A Crack In Creation Jennifer A. Doudna, Samuel H. Sternberg, 2017-06-13 BY THE WINNER OF THE 2020 NOBEL PRIZE IN CHEMISTRY | Finalist for the Los Angeles Times Book Prize “A powerful mix of science and ethics . . . This book is required reading for every concerned citizen—the material it covers should be discussed in schools, colleges, and universities throughout the country.”— New York Review of Books Not since the atomic bomb has a technology so alarmed its inventors that they warned the world about its use. That is, until 2015, when biologist Jennifer Doudna called for a worldwide moratorium on the use of the gene-editing tool CRISPR—a revolutionary new technology that she helped create—to make heritable changes in human embryos. The cheapest, simplest, most effective way of manipulating DNA ever known, CRISPR may well give us the cure to HIV, genetic diseases, and some cancers. Yet even the tiniest changes to DNA could have myriad, unforeseeable consequences, to say nothing of the ethical and societal repercussions of intentionally mutating embryos to create “better” humans. Writing with fellow researcher Sam Sternberg, Doudna—who has since won the Nobel Prize for her CRISPR research—shares the thrilling story of her discovery and describes the enormous responsibility that comes with the power to rewrite the code of life. “The future is in our hands as never before, and this book explains the stakes like no other.” — George Lucas “An invaluable account . . . We owe Doudna several times over.” — Guardian |
| crispr cas9 guide rna design: CRISPR-Cas Methods M. Tofazzal Islam, Kutubuddin Ali Molla, 2021-07-31 This second volume provides new and updated methods detailing advancements in CRISPR-Cas technical protocols. Chapters guide readers through protocols on prime editing, base editing, multiplex editing, editing in cell-free extract, in silico analysis of gRNA secondary structure and CRISPR-diagnosis. Authoritative and cutting-edge, CRISPR-Cas Methods, Volume 2 aims to serves as a laboratory manual providing scientists with a holistic view of CRISPR-Cas methodologies and its practical application for the editing of crop plants, cell lines, nematode and microorganism. The chapter “CRISPR/Cas9-mediated gene editing in human induced pluripotent stem cells” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com. |
| crispr cas9 guide rna design: Gene Correction Francesca Storici, 2016-09-03 Gene correction is a technology that gives us the tools for both repairing and mutating DNA, for discovering gene functions and for engineering new genetic variants. Gene Correction: Methods and Protocols provides a user friendly, detailed and up-to-date collection of strategies and methodologies utilized for generating specific sequence changes in the DNA of cells in the laboratory, while also tackling the major problems that the field of gene correction faces. This volume brings together many experts in the field of gene correction to disclose a wide and varied array of specific gene correction protocols for engineering mutations in DNA, for delivering correcting DNA to target cells, and for improving the accuracy and safety of the gene correction process. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Gene Correction: Methods and Protocols seeks to serve scientists of all backgrounds interested in the area of gene targeting/recombination/therapy. |
| crispr cas9 guide rna design: Genome Editing in Animals Izuho Hatada, 2023-03-11 This second edition provides new and updated protocols that can be used for generation of knockout animals. Chapters guide the reader through basic protocols for three genome editing technologies, target design tools, and specific protocols for each animal. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Genome Editing in Animals: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field. |
| crispr cas9 guide rna design: Epigenetic Reprogramming During Mouse Embryogenesis Katia Ancelin, Maud Borensztein, 2020-10-20 This volume explores the latest techniques used to study and understand chromatin reprogramming in embryos and germ cells. Various culture systems are presented, which consist of invaluable tools to investigate many developmental processes. This book also looks at methods for direct examination of DNA, RNA, and proteins in embryos, along with low-input and single-cell assays for exploring these features at the genome-wide scale. Written in the highly successful Methods in Molecular Biology 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 avoiding known pitfalls. Cutting-edge and thorough, Epigenetics Reprogramming During Mouse Embryogenesis: Methods and Protocols is a valuable resource for any scientist and researcher looking to make new discoveries in this fascinating field of chromatin reprogramming. |
| crispr cas9 guide rna design: RNA Design Alexander Churkin, |
| crispr cas9 guide rna design: CRISPR/Cas Genome Editing Anjanabha Bhattacharya, Vilas Parkhi, Bharat Char, 2020-12-11 This book offers a comprehensive collection of papers on CRISPR/Cas genome editing in connection with agriculture, climate-smart crops, food security, translational research applications, bioinformatics analysis, practical applications in cereals, floriculture crops, engineering plants for abiotic stress resistance, the intellectual landscape, regulatory framework, and policy decisions. Gathering contributions by internationally respected experts in the field of CRISPR/Cas genome editing, the book offers an essential guide for researchers, students, teachers and scientists in academia; policymakers; and public companies, private companies and cooperatives interested in understanding and/or applying CRISPR/Cas genome editing to develop new agricultural products. |
Chapter 19
Here, we summarize the current knowledge for guide RNA design and highlight discrepancies between different experimental systems. Targeted manipulation of DNA through site-specific …
Optimized CRISPR guide RNA design for two high-fidelity …
In this study, we perform a genome-scale screen in human cells to measure gRNA activity for WT-SpCas9, eSpCas9(1.1), and SpCas9-HF1 by using a high-throughput method and obtain an …
CRISPR–Cas9-mediated genome editing and guide RNA …
Here we briefly review this fast moving field, introduce the CRISPR–Cas9 system and its application to genome editing, with a focus on the basic considerations in designing the …
Research Paper crispRdesignR: A Versatile Guide RNA Design …
The success of CRISPR/Cas9 gene editing applications relies on the efficiency of the single guide RNA (sgRNA) used in conjunction with the Cas9 protein. Current sgRNA design software vary …
Crackling Cloud: an event-driven, cloud-based CRISPR-Cas9 …
Dec 4, 2024 · Gene editing has been revolutionised by the CRISPR-Cas9 tech-nology. The versatility and ease-of-use of the technology far ex-ceeds its predecessors, however, the …
Design and Validation of Guide RNAs for CRISPR Cas9 …
In an accompanying protocol (Protocol: Validating Single-Guide RNA for Aedes aegypti Gene Editing [Lo and Matthews 2023]), we detail how to design and synthesize sgRNAs as well as …
Agilent Integrated Solutions for Design, Synthesis, and Quality …
how several Agilent products can be used in a CRISPR-Cas9 workflow, starting with the design of a guide RNA (gRNA) against a specific target, synthesis of the gRNA, quality control methods …
A Guide RNA Sequence Design Platform for the CRISPR/Cas9 …
Thus, we developed a Guide RNA Sequence Design Platform for the Cas9/CRISPR silencing system for model organisms. The platform is easy to use for gRNA design with input query …
Simple protocol for gene editing using GenCrisprTM Cas9 …
Compared with traditional genetic engineering, the CRISPR-Cas9 system is easier to design and simpler to use because targeting a new locus only requires the redesign of a single guide RNA …
A CRISPR/Cas Guide RNA Design In Silico Activity
CRISPR/Cas tool design requires students to consider where the gene should be targeted, the intended consequence of the targeting, what mechanisms are available to introduce foreign
Cas9 guide-RNA Design Tutorial for Sage HLS-CATCH - Sage …
The general approach to guide-RNA (gRNA) design relies on reference sequences for human and mouse genomes and the highly featured annotations available from University of California …
CRISPR-P: A Web Tool for Synthetic Single-Guide RNA Design …
CRISPR-system in plant genome editing, tools for opti-mized sgRNA selection in plants are highly needed. In the current analysis, we present a web application tool— CRISPR-P, for CRISPR …
crisprDesign: Comprehensive design of CRISPR gRNAs for …
1.10.0 Provides a comprehensive suite of functions to design and anno-tate CRISPR guide RNA (gRNAs) sequences. This includes on- and off-target search, on-target eficiency scoring, off …
CRISPR/Cas9 gRNA Vector - GenScript
With CRISPR/Cas9 genome editing, modified clonal cell lines can be derived within 2-3 weeks starting from the gRNA design stage, while transgenic animal strains can be created in a …
Design of single guide RNA for CRISPR/Cas9 mediated …
We designed sgRNAs to target the TMS5 and SD1 gene of rice using CRISPR-PLANT, an online platform for designing guide RNAs. The efficiency of the sgRNAs were assessed for various …
Overview of guide RNA design tools for CRISPR-Cas9 genome …
Here, I provide an overview of the technology, focusing on guide RNA design principles, available software tools and their strengths and weaknesses. endonuclease is the most recent …
Rational design of minimum CRISPR guide RNA by site …
We report a Cas9–RNA conjugation strategy to afford minimal crRNA containing only the guide sequence for the target gene, which may simplify and reduce the cost for large-scale and...
CRISPR Design tool FAQ - Horizon Discovery
The CRISPR-Cas9 mechanism cuts both DNA strands. Therefore, it is possible to target a region using a guide RNA that targets either DNA strand. What is essential is ensuring the PAM is in …
CrispRdesignR: A Versatile Guide RNA Design Package in R for …
16 The CRISPR/Cas9 system has attracted attention in recent years for its ability to edit and 17 regulate DNA in a wide variety of organisms and cell types. Using a strand of single guide …
Design high specificity CRISPR-Cas9 gRNAs: principles and …
CRISPR-Cas9 is an efficient and easy to implement system for genome editing. Cas9 can tolerate 1-3 mismatches and generate off-target mutations. gRNA design steps: Target gene analysis, …
Chapter 19
Here, we summarize the current knowledge for guide RNA design and highlight discrepancies between different experimental systems. Targeted manipulation of DNA through site-specific …
Optimized CRISPR guide RNA design for two high-fidelity …
In this study, we perform a genome-scale screen in human cells to measure gRNA activity for WT-SpCas9, eSpCas9(1.1), and SpCas9-HF1 by using a high-throughput method and obtain an …
CRISPR–Cas9-mediated genome editing and guide RNA …
Here we briefly review this fast moving field, introduce the CRISPR–Cas9 system and its application to genome editing, with a focus on the basic considerations in designing the …
Research Paper crispRdesignR: A Versatile Guide RNA Design …
The success of CRISPR/Cas9 gene editing applications relies on the efficiency of the single guide RNA (sgRNA) used in conjunction with the Cas9 protein. Current sgRNA design software vary …
Crackling Cloud: an event-driven, cloud-based CRISPR-Cas9 …
Dec 4, 2024 · Gene editing has been revolutionised by the CRISPR-Cas9 tech-nology. The versatility and ease-of-use of the technology far ex-ceeds its predecessors, however, the …
Design and Validation of Guide RNAs for CRISPR Cas9 …
In an accompanying protocol (Protocol: Validating Single-Guide RNA for Aedes aegypti Gene Editing [Lo and Matthews 2023]), we detail how to design and synthesize sgRNAs as well as …
Agilent Integrated Solutions for Design, Synthesis, and …
how several Agilent products can be used in a CRISPR-Cas9 workflow, starting with the design of a guide RNA (gRNA) against a specific target, synthesis of the gRNA, quality control methods …
A Guide RNA Sequence Design Platform for the …
Thus, we developed a Guide RNA Sequence Design Platform for the Cas9/CRISPR silencing system for model organisms. The platform is easy to use for gRNA design with input query …
Simple protocol for gene editing using GenCrisprTM Cas9 …
Compared with traditional genetic engineering, the CRISPR-Cas9 system is easier to design and simpler to use because targeting a new locus only requires the redesign of a single guide RNA …
A CRISPR/Cas Guide RNA Design In Silico Activity
CRISPR/Cas tool design requires students to consider where the gene should be targeted, the intended consequence of the targeting, what mechanisms are available to introduce foreign
Cas9 guide-RNA Design Tutorial for Sage HLS-CATCH - Sage …
The general approach to guide-RNA (gRNA) design relies on reference sequences for human and mouse genomes and the highly featured annotations available from University of California …
CRISPR-P: A Web Tool for Synthetic Single-Guide RNA …
CRISPR-system in plant genome editing, tools for opti-mized sgRNA selection in plants are highly needed. In the current analysis, we present a web application tool— CRISPR-P, for CRISPR …
crisprDesign: Comprehensive design of CRISPR gRNAs for …
1.10.0 Provides a comprehensive suite of functions to design and anno-tate CRISPR guide RNA (gRNAs) sequences. This includes on- and off-target search, on-target eficiency scoring, off …
CRISPR/Cas9 gRNA Vector - GenScript
With CRISPR/Cas9 genome editing, modified clonal cell lines can be derived within 2-3 weeks starting from the gRNA design stage, while transgenic animal strains can be created in a …
Design of single guide RNA for CRISPR/Cas9 mediated …
We designed sgRNAs to target the TMS5 and SD1 gene of rice using CRISPR-PLANT, an online platform for designing guide RNAs. The efficiency of the sgRNAs were assessed for various …
Overview of guide RNA design tools for CRISPR-Cas9 …
Here, I provide an overview of the technology, focusing on guide RNA design principles, available software tools and their strengths and weaknesses. endonuclease is the most recent …
Rational design of minimum CRISPR guide RNA by site …
We report a Cas9–RNA conjugation strategy to afford minimal crRNA containing only the guide sequence for the target gene, which may simplify and reduce the cost for large-scale and...
CRISPR Design tool FAQ - Horizon Discovery
The CRISPR-Cas9 mechanism cuts both DNA strands. Therefore, it is possible to target a region using a guide RNA that targets either DNA strand. What is essential is ensuring the PAM is in …
CrispRdesignR: A Versatile Guide RNA Design Package in R …
16 The CRISPR/Cas9 system has attracted attention in recent years for its ability to edit and 17 regulate DNA in a wide variety of organisms and cell types. Using a strand of single guide …
