Crispr And Rnai Systems

Plants are vulnerable to pathogens including fungi, bacteria, and viruses, which cause critical problems and deficits. Crop protection by plant breeding delivers a promising solution with no obvious effect on human health or the local ecosystem. Crop improvement has been the most powerful approach for producing unique crop cultivars since domestication occurred, making possible the main innovations in feeding the globe and community development. Genome editing is one of the genetic devices that can be implemented, and disease resistance is frequently cited as the most encouraging application of CRISPR/Cas9 technology in agriculture. Nanobiotechnology has harnessed the power of genome editing to develop agricultural crops. Nanosized DNA or RNA nanotechnology approaches could contribute to raising the stability and performance of CRISPR guide RNAs. This book brings together the latest research in these areas. CRISPR and RNAi Systems: Nanobiotechnology Approaches to Plant Breeding and Protection presents a complete understanding of the RNAi and CRISPR/Cas9 techniques for controlling mycotoxins, fighting plant nematodes, and detecting plant pathogens. CRISPR/Cas genome editing enables efficient targeted modification in most crops, thus promising to accelerate crop improvement. CRISPR/Cas9 can be used for management of plant insects, and various plant pathogens. The book is an important reference source for both plant scientists and environmental scientists who want to understand how nano biotechnologically based approaches are being used to create more efficient plant protection and plant breeding systems. Shows how nanotechnology is being used as the basis for new solutions for more efficient plant breeding and plant protection Outlines the major techniques and applications of both CRISPR and RNAi technologies Assesses the major challenges of escalating these technologies on a mass scale

This volume explores the uses of RNAi and CRISPR interferences as a general method for inhibiting gene expression, with focus on their biological functions, design, chemical modifications, delivery, and preclinical/clinical applications. In addition to relevant backgrounds, the chapters in this book discuss simple and accurate protocols dealing with the latest advances in RNAi and CRISPR applications and look at how these methods differ from one another. 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 cutting-edge, RNA Interference and CRISPR Technologies: Technical Advances and New Therapeutic Opportunities is a valuable resource for any scientist, teachers, graduate student, postdoc, and clinician interested in this field. This book also benefits anyone in research and development in biotech and pharmaceutical companies who want to understand more about these technologies, and their applications in biology and medicine. .

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. Provides basic understanding and a clear picture on how to design, use and implement the CRISPR-Cas9 system in different organisms Explains how to create an animal model for disease research and screening purposes using CRISPR Discusses the application of CRISPR-Cas9 systems in basic sciences, biomedicine, virology, bacteriology, molecular biology, neurology, cancer, industry, and many more

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/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.

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.

Reprogramming the Genome: Applications of CRISPR-Cas in Non-mammalian Systems Part B, represents the collation of chapters written by eminent scientists worldwide. 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. It is a simple, rapid, precise, and cost-effective tool for genome editing and regulation of a wide range of organisms. It has gained scientific and public attention worldwide. This volume mainly covers insect cell line, protozoans, zebrafish, drosophila, CRISPRi, patents as well as technology transfer, and many more. This book is a key source of information available in a single volume. This book will be useful for not only beginners in genome engineering, but also students, researchers, scientists, policymakers, and stakeholders interested in harnessing the potential of reprogramming of the genomes in several areas. Offers basic understanding and a 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 insect cell line, protozoans, zebrafish, drosophila, and many more Discusses the advances, patents, applications, challenges and opportunities in CRISPR-Cas9 systems in basic sciences, biomedicine, molecular biology and many more

This book facilitates an integrative understanding of the development, genetics and evolution of butterfly wing patterns. To develop a deep and realistic understanding of the diversity and evolution of butterfly wing patterns, it is essential and necessary to approach the problem from various kinds of key research fields such as “evo-devo,” “eco-devo,” ”developmental genetics,” “ecology and adaptation,” “food plants,” and “theoretical modeling.” The past decade-and-a-half has seen a veritable revolution in our understanding of the development, genetics and evolution of butterfly wing patterns. In addition, studies of how environmental and climatic factors affect the expression of color patterns has led to increasingly deeper understanding of the pervasiveness and underlying mechanisms of phenotypic plasticity. In recognition of the great progress in research on the biology, an international meeting titled “Integrative Approach to Understanding the Diversity of Butterfly Wing Patterns (IABP-2016)” was held at Chubu University, Japan in August 2016. This book consists of selected contributions from the meeting. Authors include main active researchers of new findings of corresponding genes as well as world leaders in both experimental and theoretical approaches to wing color patterns. The book provides excellent case studies for graduate and undergraduate classes in evolution, genetics/genomics, developmental biology, ecology, biochemistry, and also theoretical biology, opening the door to a new era in the integrative approach to the analysis of biological problems. This book is open access under a CC BY 4.0 license.

Gene silencing technologies such as clustered regulatory short palindromic repeats (CRISPR)/Cas9 and RNA interference (RNAi) created a revolution in genome engineering. They are highly site-specific, simple, fast, and cost-effective. Since their discovery, gene silencing technologies have extensively been implemented in various organisms including humans, animals, plants, and microbes. They have been used for both basic science studies such as gene functional analysis and applied science such as medicine and crop improvement. In this work, we used multiplex CRISPR/Cas9 system to knock out all four members of the class II glutaredoxin (GRX) gene family including S14, S15, S16 and S17 in tomato and RNAi technology to express mouse complement 3 (C3) and complement factor 7 (CF7) small interfering RNAs (siRNAs) in lettuce. Reactive oxygen species (ROS) are induced under abiotic and biotic stresses and also as byproducts of aerobic metabolism, and their overproduction causes oxidative damage to macromolecules such as proteins, lipids, carbohydrates, and nucleic acids. GRXs are small ubiquitous proteins that are known to be involved in cellular redox homeostasis by reducing disulfate bonds and scavenging ROS. To investigate the functions of each member of class II GRX gene family in tomato's response to abiotic stresses, single and multiple knockout lines for class II GRXs were obtained using multiplex CRISPR/Cas9 system. Mutant lines and wild-type plants were subjected to heat, drought, chilling, cadmium (Cd) toxicity, and short photoperiod stresses. Phenotyping data showed that members of the class II GRX gene family are critical for tomato's growth, development, and survival under several abiotic stresses. Our findings propose novel functions for members of class II SlGRXs. RNAi technology can be utilized to target disease-related proteins. However, the application of siRNAs is challenging predominantly due to the difficult delivery and instability of siRNA into the host system. Recent findings of bioactivity and bioavailability of plants' miRNAs through animals' digestive systems led to the newly introduced field of dietary siRNAs. Animal's siRNA can be expressed in plant tissues and delivered as dietary siRNA. Here, an expression vector made based on a rice miRNA backbone, Osa-MIR528, was utilized to construct two plant expression vectors containing siRNAs silencing mouse C3 and CF7 proteins. Both C3 and CF7 proteins are involved in blood clotting which could lead to cardiovascular dysfunction. Expression of both primary and mature C3 and CF7 siRNAs in lettuce was validated by semi quantitative real-time PCR and end-point PCR, respectively, and was confirmed via Sanger sequencing. Established amiRNA system in lettuce through this work will have further applications. As an edible leafy plant with high biomass, lettuce can be used as a valuable host to produce various diseases targeting siRNAs.

RNA interference (RNAi), a hallmark of all biological sciences of twenty-first century, is an evolutionarily conserved and double-stranded RNA-dependent eukaryotic cell defense process. Opportunity to utilize an organisms own gene and to systematically induce and trigger RNAi for any desired sequence made RNAi an efficient approach for functional genomics, providing a solution for conventional longstanding obstacles in life sciences. RNAi research and application have significantly advanced during past two decades. This book RNA interference provides an updated knowledge and progress on RNAi in various organisms, explaining basic principles, types, and property of inducers, structural modifications, delivery systems/methodologies, and various successful bench-to-field or clinic applications and disease therapies with some aspects of limitations, alternative tools, safety, and risk assessment.

This two-volume set highlights the various innovative and emerging techniques and molecular applications that are currently being used in plant abiotic stress physiology. Volume 1: Responses and Adaptations focuses on the responses and adaptations of plants to stress factors at the cellular and molecular levels and offers a variety of advanced management strategies and technologies. Volume 2: Molecular Advancements introduces a range of state-of-the-art molecular advances for the mitigation of abiotic stress in plants. With contributions from specialists in the field, Volume 1 first discusses the physiology and defense mechanisms of plants and the various kinds of stress, such as from challenging environments, climate change, and nutritional deficiencies. It goes on to discuss trailblazing management techniques that include genetics approaches for improving abiotic stress tolerance in crop plants along with CRISPR/CAS-mediated genome editing technologies. Volume 2 discusses how plants have developed diverse physiological and molecular adjustments to safeguard themselves under challenging conditions and how emerging new technologies can utilize these plant adaptations to enhance plant resistance. These include using plant-environment interactions to develop crop species that are resilient to climate change, applying genomics and phenomics approaches from the study of abiotic stress tolerance and more. Agriculture today faces countless challenges to meet the rising need for sustainable food supplies and guarantees of high-quality nourishment for a quickly increasing population. To ensure sufficient food production, it is necessary to address the difficult environmental circumstances that are causing cellular oxidative stress in plants due to abiotic factors, which play a defining role in shaping yield of crop plants. These two volumes help to meet these challenges by providing a rich source of information on plant abiotic stress physiology and effective management techniques.

The Logic of Chance offers a reappraisal and a new synthesis of theories, concepts, and hypotheses on the key aspects of the evolution of life on earth in light of comparative genomics and systems biology. The author presents many specific examples from systems and comparative genomic analysis to begin to build a new, much more detailed, complex, and realistic picture of evolution. The book examines a broad range of topics in evolutionary biology including the inadequacy of natural selection and adaptation as the only or even the main mode of evolution; the key role of horizontal gene transfer in evolution and the consequent overhaul of the Tree of Life concept; the central, underappreciated evolutionary importance of viruses; the origin of eukaryotes as a result of endosymbiosis; the concomitant origin of cells and viruses on the primordial earth; universal dependences between genomic and molecular-phenomic variables; and the evolving landscape of constraints that shape the evolution of genomes and molecular phenomes. "Koonin's account of viral and pre-eukaryotic evolution is undoubtedly up-to-date. His "mega views" of evolution (given what was said above) and his cosmological musings, on the other hand, are interesting reading." Summing Up: Recommended Reprinted with permission from CHOICE, copyright by the American Library Association.

In the most recent years, each of the RNA silencing pathways of plants have appeared to generate ncRNAs with dedicated functions, specialized biological activities and specific functional scopes. RNA silencing plays a crucial role in coordinating the expression, stability, protection and inheritance of eukaryotic genomes. It compromises several mechanisms, that invariably depend on core small non coding RNAs and that achieve dedicated sequence-specific functions. RNA silencing has been recognized to carry critical developmental, stress-response and bodyguard functions be coordinating the expression, protection, stability and inheritance of virtually all eukaryotic genomes. Thus, the ncRNAs encompass a wide set of mechanisms that achieve specialized functions.

Ever since the birth of molecular biology, the tantalizing possibility of treating disease at its genetic roots has become increasingly feasible. Gene therapy - though still in its infancy - remains one of the hottest areas of research in medicine. Its approach utilizes a gene transfer vehicle ( vector) to deliver therapeutic DNA or RNA to cells of the body in order to rectify the defect that is causing the disease. Successful therapies have been reported in humans in recent years such as cures in boys with severe immune deficiencies. Moreover, gene therapy strategies are being adapted in numerous biomedical laboratories to obtain novel treatments for a variety of diseases and to study basic biological aspects of disease. Correction of disease in animal studies, is steadily gaining ground, highlighting the immense potential of gene therapy in the medical profession. This book will cover topics that are at the forefront of biomedical research such as RNA interference, viral and non-viral gene transfer systems, treatment of hematological diseases and disorders of the central nervous system.Leading experts on the respective vector or disease will contribute the individual chapters and explain cutting-edge technologies. It also gives a broad overview of the most important gene transfer vectors and most extensively studied target diseases. This comprehensive guide is therefore a must-read for anyone in the biotechnology, biomedical or medical industries seeking to further their knowledge in the area of human gene therapy.

This volume, the first of a two-part series, covers topics including historical, ecological and evolutionary considerations, genomics and molecular biology, and interaction of phages with their hosts. Contributions from leading authorities Informs and updates on all the latest developments in the field

Biotechnology, Second Edition approaches modern biotechnology from a molecular basis, which has grown out of increasing biochemical understanding of genetics and physiology. Using straightforward, less-technical jargon, Clark and Pazdernik introduce each chapter with basic concepts that develop into more specific and detailed applications. This up-to-date text covers a wide realm of topics including forensics, bioethics, and nanobiotechnology using colorful illustrations and concise applications. In addition, the book integrates recent, relevant primary research articles for each chapter, which are presented on an accompanying website. The articles demonstrate key concepts or applications of the concepts presented in the chapter, which allows the reader to see how the foundational knowledge in this textbook bridges into primary research. This book helps readers understand what molecular biotechnology actually is as a scientific discipline, how research in this area is conducted, and how this technology may impact the future. Up-to-date text focuses on modern biotechnology with a molecular foundation Includes clear, color illustrations of key topics and concept Features clearly written without overly technical jargon or complicated examples Provides a comprehensive supplements package with an easy-to-use study guide, full primary research articles that demonstrate how research is conducted, and instructor-only resources

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

During the past 15 years, cellular and molecular approaches have emerged as valuable adjuncts to supplement and complement conventional breeding methods for a wide variety of crop plants. Biotechnology increasingly plays a role in the creation, conservation, characterization and utilization of genetic variability for germplasm enhancement. For instance, anther/microspore culture, somaclonal variation, embryo culture and somatic hybridization are being exploited for obtaining incremental improvement in the existing cultivars. In addition, genes that confer insect- and disease-resistance, abiotic stress tolerance, herbicide tolerance and quality traits have been isolated and re-introduced into otherwise sensitive or susceptible species by a variety of transgenic techniques. Together these transformative methodologies grant access to a greater repertoire of genetic diversity as the gene(s) may come from viruses, bacteria, fungi, insects, animals, human beings, unrelated plants or even be artificially derived. Remarkable achievements have been made in the production, characterization, field evaluation and commercialization of transgenic crop varieties worldwide. Likewise, significant advances have been made towards increasing crop yields, improving nutritional quality, enabling crops to be raised under adverse conditions and developing resistance to pests and diseases for sustaining global food and nutritional security. The overarching purpose of this 3-volume work is to summarize the history of crop improvement from a technological perspective but to do so with a forward outlook on further advancement and adaptability to a changing world. Our carefully chosen “case studies of important plant crops” intend to serve a diverse spectrum of audience looking for the right tools to tackle complicated local and global issues.

Molecular Biology, Second Edition, examines the basic concepts of molecular biology while incorporating primary literature from today’s leading researchers. This updated edition includes Focuses on Relevant Research sections that integrate primary literature from Cell Press and focus on helping the student learn how to read and understand research to prepare them for the scientific world. The new Academic Cell Study Guide features all the articles from the text with concurrent case studies to help students build foundations in the content while allowing them to make the appropriate connections to the text. Animations provided deal with topics such as protein purification, transcription, splicing reactions, cell division and DNA replication and SDS-PAGE. The text also includes updated chapters on Genomics and Systems Biology, Proteomics, Bacterial Genetics and Molecular Evolution and RNA. An updated ancillary package includes flashcards, online self quizzing, references with links to outside content and PowerPoint slides with images. This text is designed for undergraduate students taking a course in Molecular Biology and upper-level students studying Cell Biology, Microbiology, Genetics, Biology, Pharmacology, Biotechnology, Biochemistry, and Agriculture. NEW: "Focus On Relevant Research" sections integrate primary literature from Cell Press and focus on helping the student learn how to read and understand research to prepare them for the scientific world. NEW: Academic Cell Study Guide features all articles from the text with concurrent case studies to help students build foundations in the content while allowing them to make the appropriate connections to the text. NEW: Animations provided include topics in protein purification, transcription, splicing reactions, cell division and DNA replication and SDS-PAGE Updated chapters on Genomics and Systems Biology, Proteomics, Bacterial Genetics and Molecular Evolution and RNA Updated ancillary package includes flashcards, online self quizzing, references with links to outside content and PowerPoint slides with images. Fully revised art program