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

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.

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. .

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.

This book offers a unique and comprehensive overview of key RNA-based technologies, as well as their development and applications for the functional genomics of plant coding and non-coding genes. It focuses on the latest as well as classical RNA-based techniques used for studies on small RNAs, long non-coding RNAs and protein-coding genes. These techniques chiefly focus on target mimics (TMs) and short tandem target mimics (STTMs) for small RNAs, and artificial microRNAs (amiRNAs), RNA interference (RNAi) and CRISPR/Cas for genes. Furthermore, the book discusses the latest trends in the field and various modifications of the above-mentioned approaches, and explores how these RNA-based technologies have been developed, applied and validated as essential technologies in plant functional genomics. RNA-based technologies, their mechanisms of action, their advantages and disadvantages, and insights into the further development and applications of these technologies in plants are discussed. These techniques will enable the users to functionally characterize genes and small RNAs through silencing, overexpression and editing. Gathering contributions by globally respected experts, the book will appeal to students, teachers and scientists in academia and industry who are interested in horticulture, genetics, pathology, entomology, physiology, molecular genetics and breeding, in vitro culture & genetic engineering, and functional genomics.

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.

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

The CRISPR-Cas9 genome-editing system is creating a revolution in the science world. In the laboratory, CRISPR-Cas9 can efficiently be used to target specific genes, correct mutations and regulate gene expression of a wide array of cells and organisms, including human cells. CRISPR-/Cas9 Based Genome Editing for Treating Genetic Disorders and Diseases is a unique reading material for college students, academicians, and other health professionals interested in learning about the broad range of applications of CRISPR/Cas9 genetic scissors. Some topics included in this book are: the role of the CRISPR/Cas9 system in neuroscience, gene therapy, epigenome editing, genome mapping, cancer, virus infection control strategies, regulatory challenges and bioethical considerations.

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.

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

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.

Overviews of biochemical, genetic, and molecular perspectives of plant-insect interactions with added emphasis on bioinformatic, genomic, and transcriptome analysis are comprehensively treated in this book. It presents the agro-ecological and evolutionary aspects of plant-insect interactions with an exclusive focus on the climate change effect on the resetting of plant-insect interactions. A valuable resource for biotechnologists, entomologists, agricultural scientists, and policymakers, the book includes theoretical aspects as a base toward real-world applications of holistic integrated pest management in agro-ecosystems.

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 book is open access under a CC BY 4.0 license. CRISPR-Cas9 is a rapid, efficient, versatile and relatively cheap method for dissecting the molecular pathways that are the basis of life, as well as for investigating and potentially rectifying faults in these pathways that result in disease. This book reviews how CRISPR-Cas9 and other genome editing techniques are advancing our understanding of development and function in the nervous system, uncovering the molecular causes of neurological disorders and providing tools for gene therapy.

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.

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.

This volume details methods of identifying synthetic lethal, genetic interactions by various approaches in different model systems including human cancer cells. Chapters guide readers through genetic interactions in model organisms, RNA interference, CRISPR/Cas9 based genome editing technologies, drug-gene interactions, mapping chemical genetic interactions, synergistic drug-gene relations, single cell sequencing, gene expression profiling, and novel genetic interactions. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Genetic Interaction Mapping aims to be a useful practical guide to researches to help further their study in this field.

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