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| crispr cell line engineering: 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 cell line engineering: 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 cell line engineering: Cell Culture Engineering Gyun Min Lee, Helene Faustrup Kildegaard, 2020-01-13 Offers a comprehensive overview of cell culture engineering, providing insight into cell engineering, systems biology approaches and processing technology In Cell Culture Engineering: Recombinant Protein Production, editors Gyun Min Lee and Helene Faustrup Kildegaard assemble top class authors to present expert coverage of topics such as: cell line development for therapeutic protein production; development of a transient gene expression upstream platform; and CHO synthetic biology. They provide readers with everything they need to know about enhancing product and bioprocess attributes using genome-scale models of CHO metabolism; omics data and mammalian systems biotechnology; perfusion culture; and much more. This all-new, up-to-date reference covers all of the important aspects of cell culture engineering, including cell engineering, system biology approaches, and processing technology. It describes the challenges in cell line development and cell engineering, e.g. via gene editing tools like CRISPR/Cas9 and with the aim to engineer glycosylation patterns. Furthermore, it gives an overview about synthetic biology approaches applied to cell culture engineering and elaborates the use of CHO cells as common cell line for protein production. In addition, the book discusses the most important aspects of production processes, including cell culture media, batch, fed-batch, and perfusion processes as well as process analytical technology, quality by design, and scale down models. -Covers key elements of cell culture engineering applied to the production of recombinant proteins for therapeutic use -Focuses on mammalian and animal cells to help highlight synthetic and systems biology approaches to cell culture engineering, exemplified by the widely used CHO cell line -Part of the renowned Advanced Biotechnology book series Cell Culture Engineering: Recombinant Protein Production will appeal to biotechnologists, bioengineers, life scientists, chemical engineers, and PhD students in the life sciences. |
| crispr cell line engineering: ISCN 2013 International Standing Committee on Human Cytogenetic Nomenclature, 2013 This publication extends the now classic system of human cytogenetic nomenclature prepared by an expert committee and published in collaboration with Cytogenetic and Genome Research' since 1963. Revised and finalized by the ISCN Committee and its advisors at a meeting in Seattle, Wash., in April 2012, the ISCN 2013 updates, revises and incorporates all previous human cytogenetic nomenclature recommendations into one systematically organized publication that supersedes all previous ISCN recommendations. There are several new features in ISCN 2013: an update of the microarray nomenclature, many more illustrative examples of uses of nomenclature in all sections some definitions including chromothripsis and duplication a new chapter for nomenclature that can be used for any region-specific assay. The ISCN 2013 is an indispensable reference volume for human cytogeneticists, technicians and students for the interpretation and communication of human cytogenetic nomenclature. |
| crispr cell line engineering: 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 cell line engineering: Diabetic Nephropathy Luigi Gnudi, David A. Long, 2019-11-08 This book provides a toolkit of novel research approaches for investigators to study diabetic nephropathy, including critical experimental models from the fly to the fish, cells in culture, and in vivo mammalian approaches. The collection also explores powerful techniques to image the kidney, such as traditional histological techniques as well as electron, confocal, and two-photon microscopy, pathophysiology of the diabetic kidney, and gene editing and regenerative medicine. 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, Diabetic Nephropathy: Methods and Protocols seeks to foster new research directions and inspire ideas to enhance our understanding of diabetic nephropathy and to develop treatments for this condition. |
| crispr cell line engineering: 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 cell line engineering: Genome Editing in Neurosciences Rudolf Jaenisch, Feng Zhang, Fred Gage, 2020-10-08 Innovations in molecular biology are allowing neuroscientists to study the brain with unprecedented resolution, from the level of single molecules to integrated gene circuits. Chief among these innovations is the CRISPR-Cas genome editing technology, which has the precision and scalability to tackle the complexity of the brain. This Colloque Médecine et Recherche has brought together experts from around the world that are applying genome editing to address important challenges in neuroscience, including basic biology in model organisms that has the power to reveal systems-level insight into how the nervous system develops and functions as well as research focused on understanding and treating human neurological disorders. This work was published by Saint Philip Street Press pursuant to a Creative Commons license permitting commercial use. All rights not granted by the work's license are retained by the author or authors. |
| crispr cell line engineering: ISCN International Standing Committee on Human Cytogenomic Nomenclature, 2016 The 2016 edition of the International System for Human Cytogenomic Nomenclature (ISCN 2016) offers standard nomenclature that is used to describe any genomic rearrangement identified by techniques ranging from karyotyping to FISH, microarray, various region specific assays, and DNA sequencing. Suggestions from the international cytogenetics community have been reviewed by the Standing Committee, an international group of experts, nominated by their peers. This updated edition offers: * many new examples, particularly for microarray and region specific assays * trackable changes in the main text compared to the previous edition for easier identification * a nomenclature standard to facilitate the description of chromosome rearrangements characterized by DNA sequencing developed through collaboration between the Human Genome Variation Society (HGVS) and ISCN to accommodate the increased use of sequencing technologies in the characterization of chromosomal abnormalities The ISCN 2016 is an indispensable reference volume for human cytogeneticists, molecular geneticists, technicians, and students for the interpretation and communication of human cytogenetic and molecular cytogenomic nomenclature. After a long collaboration with Cytogenetic and Genome Research, ISCN is now again a part of this leading journal on chromosome and genome research, combining the day-to-day business with the latest findings. |
| crispr cell line engineering: Cell Culture Engineering Wei-Shu Hu, 2006-08-16 Since the introduction of recombinant human growth hormone and insulin a quarter century ago, protein therapeutics has greatly broadened the ho- zon of health care. Many patients suffering with life-threatening diseases or chronic dysfunctions, which were medically untreatable not long ago, can attest to the wonder these drugs have achieved. Although the ?rst generation of p- tein therapeutics was produced in recombinant Escherichia coli, most recent products use mammalian cells as production hosts. Not long after the ?rst p- duction of recombinant proteins in E. coli, it was realized that the complex tasks of most post-translational modi?cations on proteins could only be ef?ciently carried out in mammalian cells. In the 1990s, we witnessed a rapid expansion of mammalian-cell-derived protein therapeutics, chie?y antibodies. In fact, it has been nearly a decade since the market value of mammalian-cell-derived protein therapeutics surpassed that of those produced from E. coli. A common characteristic of recent antibody products is the relatively large dose required for effective therapy, demanding larger quantities for the treatment of a given disease. This, coupled with the broadening repertoire of protein drugs, has rapidly expanded the quantity needed for clinical applications. The increasing demand for protein therapeutics has not been met exclusively by construction of new manufacturing plants and increasing total volume capacity. More - portantly the productivity of cell culture processes has been driven upward by an order of magnitude in the past decade. |
| crispr cell line engineering: Glyco-Engineering Alexandra Castilho, 2015-06-17 Conceived with the intention of providing an array of strategies and technologies currently in use for glyco-engineering distinct living organisms, this book contains a wide range of methods being developed to control the composition of carbohydrates and the properties of proteins through manipulations on the production host rather than in the protein itself. The first five sections deal with host-specific glyco-engineering and contain chapters that provide protocols for modifications of the glycosylation pathway in bacteria, yeast, insect, plants and mammalian cells, while the last two sections explore alternative approaches to host glyco-engineering and selected protocols for the analysis of the N-glycans and glyco-profiling by mass spectrometry. 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 extensive, Glyco-Engineering: Methods and Protocols offers vast options to help researchers to choose the expression system and approach that best suits their intended protein research or applications. |
| crispr cell line engineering: Cell Line Development Mohamed Al-Rubeai, 2009-08-11 Mammalian cell lines command an effective monopoly for the production of therapeutic proteins that require post-translational modifications. This unique advantage outweighs the costs associated with mammalian cell culture, which are far grater in terms of development time and manufacturing when compared to microbial culture. The development of cell lines has undergone several advances over the years, essentially to meet the requirement to cut the time and costs associated with using such a complex hosts as production platforms. This book provides a comprehensive guide to the methodology involved in the development of cell lines and the cell engineering approach that can be employed to enhance productivity, improve cell function, glycosylation and secretion and control apoptosis. It presents an overall picture of the current topics central to expression engineering including such topics as epigenetics and the use of technologies to overcome positional dependent inactivation, the use of promoter and enhancer sequences for expression of various transgenes, site directed engineering of defined chromosomal sites, and examination of the role of eukaryotic nucleus as the controller of expression of genes that are introduced for production of a desired product. It includes a review of selection methods for high producers and an application developed by a major biopharmaceutical industry to expedite the cell line development process. The potential of cell engineering approch to enhance cell lines through the manipulation of single genes that play important roles in key metabolic and regulatory pathways is also explored throughout. |
| crispr cell line engineering: 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 cell line engineering: Immune Mediators in Cancer Ivana Vancurova, Yan Zhu, 2021-01-29 This book provides a comprehensive collection of classic and cutting-edge methodologies as well as bioinformatics and genome-editing approaches that are used to quantify immune mediators and analyze their function and biological activity in cancer cells and tissues. Beginning with a section on the detection of immune mediators in samples, the volume continues with sections covering cytokine bioassays, the expression and regulation of immune mediators in cancer cells, and methods to navigate the enormous datasets created by modern DNA and RNA sequencing and proteomic technology. 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 wide-ranging, Immune Mediators in Cancer: Methods and Protocols serves as a valuable resource for biochemists, molecular biologists, cancer biologists, and immunologists, as well as for physician-scientists working in the field of immunology and cancer research. |
| crispr cell line engineering: Essentials of Glycobiology Ajit Varki, Maarten J. Chrispeels, 1999 Sugar chains (glycans) are often attached to proteins and lipids and have multiple roles in the organization and function of all organisms. Essentials of Glycobiology describes their biogenesis and function and offers a useful gateway to the understanding of glycans. |
| crispr cell line engineering: Nature's Versatile Engine: Jim Vigoreaux, 2007-06-24 Methods for Obtaining X-Ray Diffraction Patterns from Drosophila 198 Diffraction Patterns from Drosophila IFM 203 Concluding Remarks 211 Note Added in Proof 211 17. Functional and Ecological Effects of Isoform Variation in Insect Flight Muscle 214 James H. Marden Abstract 214 Introduction 215 Nature's Versatile Engine 215 The Underlying Genetics: An Underinflated Genome and a Hyperinflated Transcriptome and Proteome 216 Functional Effects of Isoform Variation 219 Alternative Splicing and the Generation of Combinatorial Complexity 220 Functional Consequences of Naturally Occurring Isoform Variation 220 18. Muscle Systems Design and Integration 230 Fritz- OlafLehmann Abstract 230 Power Requirements for Flight 230 Power Reduction 233 Power Constraints on Steering Capacity 234 Balancing Power and Control 236 Changes in Muscle Efficiency in Vivo 238 Concluding Remarks 239 From the Inside Out 19. Molecular Assays for Acto-Myosin Interactions 242 John C. Sparrow and Michael A. Geeves Abstract 242 Introduction 242 Myosin Purification and Preparation of the SI Fragment 243 Purification of Flight Muscle Actin 244 Assays of Myosin and Acto-Myosin 244 Major Conclusions Relating to the Enzymatic Properties of Insect Flight Muscle Acto-Myosin 247 Major Questions about Insect Flight Muscle Acto-Myosin Kinetics That Remain 249 20. |
| crispr cell line engineering: Modulating Gene Expression Aditi Singh, Mohammad W. Khan, 2019-05-29 RNA interference (RNAi) is a widely used technology for gene silencing and has become a key tool in a myriad of research and lead discoveries. In recent years, the mechanism of RNAi agents has been well investigated, and the technique has been optimized for better effectiveness and safety. On the other hand, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9/gRNA system is a recent, novel, targeted genome-editing technique derived from the bacterial immune system. Recent advances in gene-editing research and technologies have enabled the CRISPR Cas9 system to become a popular tool for sequence-specific gene editing to correct and modify eukaryotic systems. In this book, we will focus on the mechanisms, applications, regulations (their pros and cons), and various ways in which RNAi-based methods and CRIPSR-Cas9 technology have stimulated the modulation of gene expression, thereby making them a promising therapeutic tool to treat and prevent complex diseases and disorders. |
| crispr cell line engineering: Cell Culture Engineering Gyun Min Lee, Helene Faustrup Kildegaard, 2020-01-13 Offers a comprehensive overview of cell culture engineering, providing insight into cell engineering, systems biology approaches and processing technology In Cell Culture Engineering: Recombinant Protein Production, editors Gyun Min Lee and Helene Faustrup Kildegaard assemble top class authors to present expert coverage of topics such as: cell line development for therapeutic protein production; development of a transient gene expression upstream platform; and CHO synthetic biology. They provide readers with everything they need to know about enhancing product and bioprocess attributes using genome-scale models of CHO metabolism; omics data and mammalian systems biotechnology; perfusion culture; and much more. This all-new, up-to-date reference covers all of the important aspects of cell culture engineering, including cell engineering, system biology approaches, and processing technology. It describes the challenges in cell line development and cell engineering, e.g. via gene editing tools like CRISPR/Cas9 and with the aim to engineer glycosylation patterns. Furthermore, it gives an overview about synthetic biology approaches applied to cell culture engineering and elaborates the use of CHO cells as common cell line for protein production. In addition, the book discusses the most important aspects of production processes, including cell culture media, batch, fed-batch, and perfusion processes as well as process analytical technology, quality by design, and scale down models. -Covers key elements of cell culture engineering applied to the production of recombinant proteins for therapeutic use -Focuses on mammalian and animal cells to help highlight synthetic and systems biology approaches to cell culture engineering, exemplified by the widely used CHO cell line -Part of the renowned Advanced Biotechnology book series Cell Culture Engineering: Recombinant Protein Production will appeal to biotechnologists, bioengineers, life scientists, chemical engineers, and PhD students in the life sciences. |
| crispr cell line engineering: 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 cell line engineering: 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 cell line engineering: 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 cell line engineering: Genetically Engineered Crops National Academies of Sciences, Engineering, and Medicine, Division on Earth and Life Studies, Board on Agriculture and Natural Resources, Committee on Genetically Engineered Crops: Past Experience and Future Prospects, 2017-01-28 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. |
| crispr cell line engineering: 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 cell line engineering: The Code Breaker Walter Isaacson, 2021-03-09 A Best Book of 2021 by Bloomberg BusinessWeek, Time, and The Washington Post The bestselling author of Leonardo da Vinci and Steve Jobs returns with a “compelling” (The Washington Post) account of how Nobel Prize winner Jennifer Doudna and her colleagues launched a revolution that will allow us to cure diseases, fend off viruses, and have healthier babies. When Jennifer Doudna was in sixth grade, she came home one day to find that her dad had left a paperback titled The Double Helix on her bed. She put it aside, thinking it was one of those detective tales she loved. When she read it on a rainy Saturday, she discovered she was right, in a way. As she sped through the pages, she became enthralled by the intense drama behind the competition to discover the code of life. Even though her high school counselor told her girls didn’t become scientists, she decided she would. Driven by a passion to understand how nature works and to turn discoveries into inventions, she would help to make what the book’s author, James Watson, told her was the most important biological advance since his codiscovery of the structure of DNA. She and her collaborators turned a curiosity of nature into an invention that will transform the human race: an easy-to-use tool that can edit DNA. Known as CRISPR, it opened a brave new world of medical miracles and moral questions. The development of CRISPR and the race to create vaccines for coronavirus will hasten our transition to the next great innovation revolution. The past half-century has been a digital age, based on the microchip, computer, and internet. Now we are entering a life-science revolution. Children who study digital coding will be joined by those who study genetic code. Should we use our new evolution-hacking powers to make us less susceptible to viruses? What a wonderful boon that would be! And what about preventing depression? Hmmm…Should we allow parents, if they can afford it, to enhance the height or muscles or IQ of their kids? After helping to discover CRISPR, Doudna became a leader in wrestling with these moral issues and, with her collaborator Emmanuelle Charpentier, won the Nobel Prize in 2020. Her story is an “enthralling detective story” (Oprah Daily) that involves the most profound wonders of nature, from the origins of life to the future of our species. |
| crispr cell line engineering: 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 cell line engineering: Artificial Nucleases Marina A. Zenkova, 2004-01-07 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. |
| crispr cell line engineering: 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 cell line engineering: 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 cell line engineering: Melanoma Kristian M. Hargadon, 2022-03-26 This extensive book brings together leading melanoma researchers from across the world and highlights many of the cutting-edge protocols and experimental systems currently being used to investigate questions surrounding this disease. The volume opens with sections on 2D and 3D cell culture-based approaches for studying melanoma biology, and continues with collections of chapters examining various approaches for detecting, isolating, and characterizing circulating melanoma cells, circulating tumor DNA, and exosomes, as well as experimental procedures for studying and detecting melanoma metastasis in both pre-clinical and clinical settings, bioinformatics-based approaches, protocols for quantifying and characterizing immune cell infiltrates in both melanoma tumors and tertiary lymphoid structures, and development and evaluation of therapeutic strategies for melanoma treatment. 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 comprehensive, Melanoma: Methods and Protocols aims to serve basic research scientists and clinicians who bring questions from the clinic into the lab in order to translate observations in the laboratory into improved patient care for this highly malignant form of cancer. Chapter 14 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com. |
| crispr cell line engineering: 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 cell line engineering: Safety of Genetically Engineered Foods National Research Council, Institute of Medicine, Board on Agriculture and Natural Resources, Food and Nutrition Board, Board on Life Sciences, Committee on Identifying and Assessing Unintended Effects of Genetically Engineered Foods on Human Health, 2004-07-08 Assists policymakers in evaluating the appropriate scientific methods for detecting unintended changes in food and assessing the potential for adverse health effects from genetically modified products. In this book, the committee recommended that greater scrutiny should be given to foods containing new compounds or unusual amounts of naturally occurring substances, regardless of the method used to create them. The book offers a framework to guide federal agencies in selecting the route of safety assessment. It identifies and recommends several pre- and post-market approaches to guide the assessment of unintended compositional changes that could result from genetically modified foods and research avenues to fill the knowledge gaps. |
| crispr cell line engineering: 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 cell line engineering: 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 cell line engineering: She Has Her Mother's Laugh Carl Zimmer, 2018-05-29 2019 PEN/E.O. Wilson Literary Science Writing Award Finalist Science book of the year—The Guardian One of New York Times 100 Notable Books for 2018 One of Publishers Weekly's Top Ten Books of 2018 One of Kirkus's Best Books of 2018 One of Mental Floss's Best Books of 2018 One of Science Friday's Best Science Books of 2018 “Extraordinary”—New York Times Book Review Magisterial—The Atlantic Engrossing—Wired Leading contender as the most outstanding nonfiction work of the year—Minneapolis Star-Tribune Celebrated New York Times columnist and science writer Carl Zimmer presents a profoundly original perspective on what we pass along from generation to generation. Charles Darwin played a crucial part in turning heredity into a scientific question, and yet he failed spectacularly to answer it. The birth of genetics in the early 1900s seemed to do precisely that. Gradually, people translated their old notions about heredity into a language of genes. As the technology for studying genes became cheaper, millions of people ordered genetic tests to link themselves to missing parents, to distant ancestors, to ethnic identities... But, Zimmer writes, “Each of us carries an amalgam of fragments of DNA, stitched together from some of our many ancestors. Each piece has its own ancestry, traveling a different path back through human history. A particular fragment may sometimes be cause for worry, but most of our DNA influences who we are—our appearance, our height, our penchants—in inconceivably subtle ways.” Heredity isn’t just about genes that pass from parent to child. Heredity continues within our own bodies, as a single cell gives rise to trillions of cells that make up our bodies. We say we inherit genes from our ancestors—using a word that once referred to kingdoms and estates—but we inherit other things that matter as much or more to our lives, from microbes to technologies we use to make life more comfortable. We need a new definition of what heredity is and, through Carl Zimmer’s lucid exposition and storytelling, this resounding tour de force delivers it. Weaving historical and current scientific research, his own experience with his two daughters, and the kind of original reporting expected of one of the world’s best science journalists, Zimmer ultimately unpacks urgent bioethical quandaries arising from new biomedical technologies, but also long-standing presumptions about who we really are and what we can pass on to future generations. |
| crispr cell line engineering: Genome Stability Igor Kovalchuk, Olga Kovalchuk, 2021-07-17 Genome Stability: From Virus to Human Application, Second Edition, a volume in the Translational Epigenetics series, explores how various species maintain genome stability and genome diversification in response to environmental factors. Here, across thirty-eight chapters, leading researchers provide a deep analysis of genome stability in DNA/RNA viruses, prokaryotes, single cell eukaryotes, lower multicellular eukaryotes, and mammals, examining how epigenetic factors contribute to genome stability and how these species pass memories of encounters to progeny. Topics also include major DNA repair mechanisms, the role of chromatin in genome stability, human diseases associated with genome instability, and genome stability in response to aging. This second edition has been fully revised to address evolving research trends, including CRISPRs/Cas9 genome editing; conventional versus transgenic genome instability; breeding and genetic diseases associated with abnormal DNA repair; RNA and extrachromosomal DNA; cloning, stem cells, and embryo development; programmed genome instability; and conserved and divergent features of repair. This volume is an essential resource for geneticists, epigeneticists, and molecular biologists who are looking to gain a deeper understanding of this rapidly expanding field, and can also be of great use to advanced students who are looking to gain additional expertise in genome stability. - A deep analysis of genome stability research from various kingdoms, including epigenetics and transgenerational effects - Provides comprehensive coverage of mechanisms utilized by different organisms to maintain genomic stability - Contains applications of genome instability research and outcomes for human disease - Features all-new chapters on evolving areas of genome stability research, including CRISPRs/Cas9 genome editing, RNA and extrachromosomal DNA, programmed genome instability, and conserved and divergent features of repair |
| crispr cell line engineering: 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 cell line engineering: The Use of CRISPR/Cas9, ZFNs, and TALENs in Generating Site-specific Genome Alterations Jennifer A. Doudna, Erik J. Sontheimer, 2014 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 |
| crispr cell line engineering: Systems Genetics Florian Markowetz, Michael Boutros, 2015-07-02 Whereas genetic studies have traditionally focused on explaining heritance of single traits and their phenotypes, recent technological advances have made it possible to comprehensively dissect the genetic architecture of complex traits and quantify how genes interact to shape phenotypes. This exciting new area has been termed systems genetics and is born out of a synthesis of multiple fields, integrating a range of approaches and exploiting our increased ability to obtain quantitative and detailed measurements on a broad spectrum of phenotypes. Gathering the contributions of leading scientists, both computational and experimental, this book shows how experimental perturbations can help us to understand the link between genotype and phenotype. A snapshot of current research activity and state-of-the-art approaches to systems genetics are provided, including work from model organisms such as Saccharomyces cerevisiae and Drosophila melanogaster, as well as from human studies. |
| crispr cell line engineering: A Crack in Creation Jennifer Doudna, Samuel Sternberg, 2018-06-14 A handful of discoveries have changed the course of human history. This book is about the most recent and potentially the most powerful and dangerous of them all. It is an invention that allows us to rewrite the genetic code that shapes and controls all living beings with astonishing accuracy and ease. Thanks to it, the dreams of genetic manipulation have become a stark reality: the power to cure disease and alleviate suffering, to create new sources of food and energy, as well as to re-design any species, including humans, for our own ends. Jennifer Doudna is the co-inventor of this technology - known as CRISPR - and a scientist of worldwide renown. Writing with fellow researcher Samuel Sternberg, here she provides the definitive account of her discovery, explaining how this wondrous invention works and what it is capable of. She also asks us to consider what our new-found power means: how do we enjoy its unprecedented benefits while avoiding its equally unprecedented dangers? The future of humankind - and of all life on Earth - is at stake. This book is an essential guide to the path that now lies ahead. |
| crispr cell line engineering: The Python Language Reference Manual Guido Van Rossum, Fred L. Jr. Drake, 2011-03-01 This is a printed edition of the official Python language reference manual from the Python 3.2 distribution. It describes the syntax of Python 3 and its built-in datatypes and operators. Python is an interpreted object-oriented programming language, suitable for rapid application development and scripting. This manual is intended for advanced users who need a complete description of the Python 3 language syntax and object system. A simpler tutorial suitable for new users of Python is available in the companion volume An Introduction to Python (for Python version 3.2) (ISBN 978-1-906966-13-3). For each copy of this manual sold USD 1 is donated to the Python Software Foundation by the publisher, Network Theory Ltd. |
CRISPR - Wikipedia
CRISPR (/ ˈkrɪspər /; acronym of clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found in the genomes of prokaryotic organisms such as bacteria and …
What is CRISPR? A bioengineer explains | Stanford Report
Jun 10, 2024 · The short answer: CRISPR is an immune system used by microbes to find and eliminate unwanted invaders. Qi: CRISPR stands for “clustered interspaced short palindromic …
CRISPR | Definition, Gene Editing, Technology, Uses, & Ethics
2 days ago · CRISPR, short palindromic repeating sequences of DNA, found in most bacterial genomes, that are interrupted by so-called spacer elements, or spacers—sequences of …
CRISPR - National Human Genome Research Institute
2 days ago · CRISPR (short for “clustered regularly interspaced short palindromic repeats”) is a technology that research scientists use to selectively modify the DNA of living organisms. …
What Is CRISPR Gene Editing and How Does It Work?
Apr 25, 2023 · CRISPR is a gene editing strategy that can be used to recognize, remove and potentially change genes that cause diseases.
What is CRISPR? - New Scientist
CRISPR is a technology that can be used to edit genes and, as such, will likely change the world. The essence of CRISPR is simple: it’s a way of finding a specific bit of DNA inside a cell....
What is CRISPR/Cas9? - PMC
CRISPR/Cas9 is a gene-editing technology which involves two essential components: a guide RNA to match a desired target gene, and Cas9 (CRISPR-associated protein 9)—an …
What is CRISPR? Understanding the Revolutionary Gene Editing …
Apr 17, 2025 · CRISPR is reshaping the landscape of genetics, medicine, agriculture, and beyond, offering unprecedented control over the genetic code that defines life itself. To truly …
CRISPR: A Biotech Breakthrough - NSF - National Science …
What is CRISPR? CRISPR (short for "clustered regularly interspaced short palindromic repeats") is a natural defense mechanism found in bacteria that helps them fight off viruses. When a …
CRISPR Guide - Addgene
This guide will provide a basic understanding of CRISPR biology, introduce the various applications of CRISPR, and help you get started using CRISPR in your own research.
CRISPR - Wikipedia
CRISPR (/ ˈkrɪspər /; acronym of clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found in the genomes of prokaryotic organisms such as bacteria and …
What is CRISPR? A bioengineer explains | Stanford Report
Jun 10, 2024 · The short answer: CRISPR is an immune system used by microbes to find and eliminate unwanted invaders. Qi: CRISPR stands for “clustered interspaced short palindromic …
CRISPR | Definition, Gene Editing, Technology, Uses, & Ethics
2 days ago · CRISPR, short palindromic repeating sequences of DNA, found in most bacterial genomes, that are interrupted by so-called spacer elements, or spacers—sequences of …
CRISPR - National Human Genome Research Institute
2 days ago · CRISPR (short for “clustered regularly interspaced short palindromic repeats”) is a technology that research scientists use to selectively modify the DNA of living organisms. …
What Is CRISPR Gene Editing and How Does It Work?
Apr 25, 2023 · CRISPR is a gene editing strategy that can be used to recognize, remove and potentially change genes that cause diseases.
What is CRISPR? - New Scientist
CRISPR is a technology that can be used to edit genes and, as such, will likely change the world. The essence of CRISPR is simple: it’s a way of finding a specific bit of DNA inside a cell....
What is CRISPR/Cas9? - PMC
CRISPR/Cas9 is a gene-editing technology which involves two essential components: a guide RNA to match a desired target gene, and Cas9 (CRISPR-associated protein 9)—an …
What is CRISPR? Understanding the Revolutionary Gene Editing …
Apr 17, 2025 · CRISPR is reshaping the landscape of genetics, medicine, agriculture, and beyond, offering unprecedented control over the genetic code that defines life itself. To truly …
CRISPR: A Biotech Breakthrough - NSF - National Science …
What is CRISPR? CRISPR (short for "clustered regularly interspaced short palindromic repeats") is a natural defense mechanism found in bacteria that helps them fight off viruses. When a …
CRISPR Guide - Addgene
This guide will provide a basic understanding of CRISPR biology, introduce the various applications of CRISPR, and help you get started using CRISPR in your own research.
