The Crispr Cas Tool Kit For Genome Editing

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

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.

Zero to Genetic Engineering Hero is made to provide you with a first glimpse of the inner-workings of a cell. It further focuses on skill-building for genetic engineering and the Biology-as-a-Technology mindset (BAAT). This book is designed and written for hands-on learners who have little knowledge of biology or genetic engineering. This book focuses on the reader mastering the necessary skills of genetic engineering while learning about cells and how they function. The goal of this book is to take you from no prior biology and genetic engineering knowledge toward a basic understanding of how a cell functions, and how they are engineered, all while building the skills needed to do so.

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

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 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/Cas has emerged, as a powerful genetic engineering tool. It is simple yet highly affordable and robust. This book provides a basic introduction to the mechanism and architecture of the CRISPR/Cas. It enlightens the readers about the new advancements such as gene insertion, transcriptional activation, suppression, gene tagging, multiplexing, base editing, prime editing, and tissue culture free editing which makes it easier for crop improvement. With such technologies, application of genome editing has opened new paths for crop improvements and sustainable agriculture. The current book will be useful for beginners to understand these tools and how to use them in a better and more efficient way.

Natively functioning Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) system is a prokaryotic adaptive immune system that confers resistance to foreign genetic elements including plasmids and phages. Very recently, a two-component CRISPR-Cas technology from Streptococcus Pyogenes comprising of the RNA-guided DNA endonuclease Cas9 and the guide RNA (gRNA) has been demonstrated to enable unprecedented genome editing efficiency across all domains of life. Current applications however, employ CRISPR/Cas technology in a stand-alone fashion, isolated from the rich biological machinery of the host environment in which it is applied. In this thesis, I present a toolkit designed by integrating CRISPR/Cas technology with a wide array of mammalian molecular components, thereby enabling altogether novel applications while enhancing the efficiency of current applications. By integrating a catalytically dead version of the CRISPR/Cas protein Cas9 (dCas9) with mammalian transcriptional activator VP64 and mammalian transcriptional repressor KRAB, we build and characterize tunable, multifunctional and orthogonal CRISPR/Cas transcription factors (CRISPR-TFs) in human cells. By integrating CRISPR-TFs and Cas6/Csy4 based RNA processing with multiple mammalian RNA regulatory strategies including RNA Polymerase II (RNAP II) promoters, RNAtriple- helix structures, introns, microRNAs and ribozymes, we demonstrate efficient modulation of endogenous promoters and the implementation of tunable synthetic circuits such as multistage cascades and RNA-dependent networks that can be rewired with Csy4. In summary, our integrated toolkit enables efficient and multiplexed modulation of endogenous gene networks, construction of highly scalable and tunable synthetic gene circuits. Our toolkit can be used for perturbing endogenous networks towards developmental, therapeutic and synthetic biology applications.

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 serves the teachers, researchers and the students as a handy and concise reference as well as guidebook while designing and planning for use of the advanced technologies for crop improvement. The content of the book is designed to cover the latest genome engineering techniques for crop improvement. The conventional breeding has got its limitations such as non-availability of desired genes within the genepool. In many cases, breeding has been highly used and it has nearly reached its highest limit so far as the productivity and production of crops are concerned. However, with increasing need of food and decreasing resources, including water, land, labour, etc., to feed the growing population, the alternative available ways of increasing crop productivity need to be explored and exploited. Genome engineering has a wide scope that includes technologies such as genetic engineering and transgenesis, RNA technologies, CRISPR, cisgenics and subgenics for better productivity and more efficient biotic and abiotic stress management. Therefore, the book is planned to enlighten the readers with the advanced technologies with examples and case studies, whenever possible. Efforts will be made to emphasize on general efforts on various major food crops; however, it would also be made clear that such efforts could be taken as proofs of concepts and that this could be extrapolated keeping the demand in mind.