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.

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.

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-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

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

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.

The current advances in sequencing, data mining, DNA synthesis, cloning, in silico modeling, and genome editing have opened a new field of research known as Synthetic Genomics. The main goal of this emerging area is to engineer entire synthetic genomes from scratch using pre-designed building blocks obtained by chemical synthesis and rational design. This has opened the possibility to further improve our understanding of genome fundamentals by considering the effect of the whole biological system on biological function. Moreover, the construction of non-natural biological systems has allowed us to explore novel biological functions so far not discovered in nature. This book summarizes the current state of Synthetic Genomics, providing relevant examples in this emerging field.

This volume provides readers with wide-ranging coverage of CRISPR systems and their applications in various plant species. The chapters in this book discuss topics such as plant DNA repair and genome editing; analysis of CRISPR-induced mutations; multiplexed CRISPR/Cas9 systems; CRISPR-Cas12a (Cpf1) editing systems; and non-agrobacterium based CRISPR delivery systems. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and thorough, Plant Genome Editing with CRISPR Systems: Methods and Protocols is a valuable resource for any researcher interested in learning about and using CRISPR systems in plants.

Plant diseases are destructive and threaten virtually any crop grown on a commercial scale. They are kept in check by plant breeding strategies that have introgressed disease resistance genes into many important crops, and by the deployment of costly control measures, such as antibiotics and fungicides. However, the capacity for the agents of plant disease - viruses, bacteria, fungi, and oomycetes - to adapt to new conditions, overcoming disease resistance and becoming resistant to pesticides, is very great. For these reasons, understanding the biology of plant diseases is essential for the development of durable control strategies. Plant-Pathogen Interactions provides and overview of our current knowledge of plant-pathogen interactions and the establishment of plant disease, drawing together fundamental new information on plant infection mechanisms and host responses. The role of molecular signals, gene regulation, and the physiology of pathogenic organisms are emphasized, but the role of the prevailing environment in the conditioning of disease is also discussed. Emphasizing the broader understanding that has emerged from the use of molecular genetics and genomics, Plant-Pathogen Interactions highlights those interactions that have been most widely studied and those in which genome information has provided a new level of understanding.