Download or read book Programming Bacterial Gene Expression Using Synthetic CRISPR Cas Transcriptional Regulators written by Chen Dong and published by . This book was released on 2019 with total page 211 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bacteria play a central role in biosynthesis to produce value-added organic chemicals due to its diverse carbon and energy source preferences. Implementing synthetic transcriptional regulation devices can advance our ability to modify gene expression in bacteria for engineering production strains. The CRISPR-Cas activation (CRISPRa) system, a programmable transcriptional activator with wide applications in eukaryotic organisms, has been under-utilized in bacterial due to the lack of efficient transcriptional activation domains. This work describes our contribution to the development and understanding of bacterial CRISPR-Cas-based transcriptional regulation devices. We screened novel bacterial activation domains to be used as CRISPR-Cas activators in E. coli, and optimized our strongest activation domain, SoxS into a programmable CRISPR activator. In addition, we investigated the properties of the well-established CRISPRi repression system and found that partial repression can be achieved by controlling the expression level of the sgRNA. Combining CRISPRa and CRISPRi, we demonstrated inducible simultaneous up- and down-regulation of a dual reporter gene and the CRISPR-mediated regulation of the ethanol bioproduction pathway. Moreover, we also learned important properties of the bacterial CRISPRa system which is much more restrictive than the eukaryotic CRISPRa system. CRISPRa activity is dependent on the sigma factor that the promoter recruits, the baseline strength of the promoter, the sequence composition of the promoter, the presence of nearby transcription factors, and the precise positioning of the scRNA target. We attempted to relieve these restrictions by implementing an engineered Cas protein that can bind to a wider range of targets. Lastly, we describe our efforts to transport the CRISPRa system into other non-E.coli bacteria. CRISPR-SoxS activator proved to be active in Pseudomonas putida, but not in the other organisms we tested. Therefore, we propose to characterize host-specific activation domains for bacteria whose cellular machineries are incompatible with SoxS. Together, this work provided a novel programmable gene activation device in bacteria and sets up the foundation for the development of complex, broad-host-range bacterial cellular devices for biosynthetic applications.
Download or read book The CRISPR Cas System written by Muhammad Jamal and published by Caister Academic Press Limited. This book was released on 2017 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book CRISPR Cas Activation for Regulating Multi gene Expression in Bacteria written by Jason Fontana and published by . This book was released on 2020 with total page 196 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bacterial metabolism is comprised of large and complex gene networks that can produce valuable chemical products. In principle, these networks can be reengineered using synthetic multi-gene transcriptional programs to optimize production of high-value compounds. However, our limited ability to regulate multi-gene expression with precision makes this goal difficult to achieve. While programmable gene repression in bacteria is well-established, there is a lack of robust tools for programmable gene activation. This work describes our efforts to develop new tools for bacteria to independently target and predictably manipulate the expression levels of multiple genes. We developed a programmable CRISPR-Cas transcriptional activation (CRISPRa) system for E. coli that uses modified guide RNAs to recruit a transcriptional activator. When used in concert with CRISPRi, our tools can be used to simultaneously activate and repress multiple genes. However, to expand our ability to regulate synthetic and endogenous pathways, we need predictive rules for selecting effective target sites for CRISPRa. To uncover these rules, we systematically investigated the requirements for CRISPRa in bacteria. In contrast to comparable systems in eukaryotes, we show that bacterial CRISPRa is sensitive to the geometry of the interaction between the CRISPRa complex and the synthetic promoter. Last, we discuss engineering dynamically regulated CRISPR-Cas systems to control the timing of the expression of multiple genes. These next-generation tools will enable us to engineer sophisticated multi-gene transcriptional networks that more closely resemble natural gene networks. Together, our work provides a foundation toward rapidly discovering and implementing new multi-gene programs that improve production of high-value chemicals.
Download or read book RNA Nanotechnology written by Bin Wang and published by CRC Press. This book was released on 2014-04-02 with total page 468 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Regulation of Gene Expression by Small RNAs written by Rajesh K. Gaur and published by CRC Press. This book was released on 2009-04-27 with total page 440 pages. Available in PDF, EPUB and Kindle. Book excerpt: New Findings Revolutionize Concepts of Gene FunctionEndogenous small RNAs have been found in various organisms, including humans, mice, flies, worms, fungi, and bacteria. Furthermore, it's been shown that microRNAs acting as cellular rheostats have the ability to modulate gene expression. In higher eukaryotes, microRNAs may regulate as much as 50 p
Download or read book Signals Switches Regulons and Cascades written by Society for General Microbiology. Symposium and published by Cambridge University Press. This book was released on 2002-04-18 with total page 308 pages. Available in PDF, EPUB and Kindle. Book excerpt: Publisher Description
Download or read book CRISPR Cas Systems written by Rodolphe Barrangou and published by Springer Science & Business Media. This book was released on 2012-12-13 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Cyanobacteria Biotechnology written by Paul Hudson and published by John Wiley & Sons. This book was released on 2021-04-20 with total page 560 pages. Available in PDF, EPUB and Kindle. Book excerpt: Unites a biological and a biotechnological perspective on cyanobacteria, and includes the industrial aspects and applications of cyanobacteria Cyanobacteria Biotechnology offers a guide to the interesting and useful features of cyanobacteria metabolism that keeps true to a biotechnology vision. In one volume the book brings together both biology and biotechnology to illuminate the core acpects and principles of cyanobacteria metabolism. Designed to offer a practical approach to the metabolic engineering of cyanobacteria, the book contains relevant examples of how this metabolic "module" is currently being engineered and how it could be engineered in the future. The author includes information on the requirements and real-world experiences of the industrial applications of cyanobacteria. This important book: Brings together biology and biotechnology in order to gain insight into the industrial relevant topic of cyanobacteria Introduces the key aspects of the metabolism of cyanobacteria Presents a grounded, practical approach to the metabolic engineering of cyanobacteria Offers an analysis of the requirements and experiences for industrial cyanobacteria Provides a framework for readers to design their own processes Written for biotechnologists, microbiologists, biologists, biochemists, Cyanobacteria Biotechnology provides a systematic and clear volume that brings together the biological and biotechnological perspective on cyanobacteria.
Download or read book Synthetic Biology of Cyanobacteria written by Weiwen Zhang and published by Springer. This book was released on 2018-08-08 with total page 357 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume highlights recent breakthroughs in the interdisciplinary areas of synthetic biology, metabolic engineering and bioprocess engineering for the production of green chemicals. It also presents practical experimental and computational tools for the design, construction and manipulation of cyanobacteria cell factories. The respective contributions cover new technologies in the field, such as novel genetic transformation techniques and bioinformatics analysis methods and address various aspects of cyanobacterial synthetic biology, offering a valuable resource for students and researchers in the fields of industry microbiology and biomedical engineering.
Download or read book Synthetic Biology written by Christina Smolke and published by John Wiley & Sons. This book was released on 2018-02-28 with total page 532 pages. Available in PDF, EPUB and Kindle. Book excerpt: A review of the interdisciplinary field of synthetic biology, from genome design to spatial engineering. Written by an international panel of experts, Synthetic Biology draws from various areas of research in biology and engineering and explores the current applications to provide an authoritative overview of this burgeoning field. The text reviews the synthesis of DNA and genome engineering and offers a discussion of the parts and devices that control protein expression and activity. The authors include information on the devices that support spatial engineering, RNA switches and explore the early applications of synthetic biology in protein synthesis, generation of pathway libraries, and immunotherapy. Filled with the most recent research, compelling discussions, and unique perspectives, Synthetic Biology offers an important resource for understanding how this new branch of science can improve on applications for industry or biological research.
Download or read book Cell Free Synthetic Biology written by Jian Li and published by Frontiers Media SA. This book was released on 2022-01-13 with total page 205 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book CRISPR Cas System in Translational Biotechnology written by Swati Joshi and published by Elsevier. This book was released on 2023-11-16 with total page 317 pages. Available in PDF, EPUB and Kindle. Book excerpt: CRISPR-Cas System in Translational Biotechnology discusses applied and translational aspects of the CRISPR-Cas technology. The book bridges the gap between theoretical knowledge and practical solutions surrounding this emerging and impactful technology in several academic and industrial fields. It is split in five sections: CRISP-Cas fundamentals and advancements; CRISP-Cas in medical biotechnology; CRISP-Cas in environmental biotechnology; CRISP-Cas in food biotechnology; and biosafety, patents and commercialization of CRISP-Cas technology. Written by experts from diverse backgrounds, the content covers the subject and its impact in multiple fields. It is a valuable resource for graduate students and researchers on bioinformatics, systems biology, and members of the biomedical field and biotechnology industry who are interested in learning more about CRISP-Cas system and its applications. - Discusses applied aspects of CRISPR-Cas technology and state-of-the-art technological/translational advancements in the field - Focuses on the CRISPR-Cas mediated genetic engineering for employment in various industries such as medical, agricultural, environmental and food - Encompasses knowledge on CRISPR-Cas commercialization, potential markets and associated ethical challenges
Download or read book Synthetic Genomics written by Miguel Fernández-Niño and published by BoD – Books on Demand. This book was released on 2022-02-02 with total page 106 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Regulation of Bacterial Virulence written by Michael L. Vasil and published by American Society for Microbiology Press. This book was released on 2012-12-05 with total page 1189 pages. Available in PDF, EPUB and Kindle. Book excerpt: A comprehensive compendium of scholarly contributions relating to bacterial virulence gene regulation. • Provides insights into global control and the switch between distinct infectious states (e.g., acute vs. chronic). • Considers key issues about the mechanisms of gene regulation relating to: surface factors, exported toxins and export mechanisms. • Reflects on how the regulation of intracellular lifestyles and the response to stress can ultimately have an impact on the outcome of an infection. • Highlights and examines some emerging regulatory mechanisms of special significance. • Serves as an ideal compendium of valuable topics for students, researchers and faculty with interests in how the mechanisms of gene regulation ultimately affect the outcome of an array of bacterial infectious diseases.
Download or read book Mammalian Cell Engineering written by Ryosuke Kojima and published by Humana. This book was released on 2022-07-21 with total page 330 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Genetically Engineered Crops written by National Academies of Sciences, Engineering, and Medicine and published by National Academies Press. This book was released on 2017-01-28 with total page 607 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book The Chemistry of Microbiomes written by National Academies of Sciences, Engineering, and Medicine and published by National Academies Press. This book was released on 2017-07-19 with total page 133 pages. Available in PDF, EPUB and Kindle. Book excerpt: The 21st century has witnessed a complete revolution in the understanding and description of bacteria in eco- systems and microbial assemblages, and how they are regulated by complex interactions among microbes, hosts, and environments. The human organism is no longer considered a monolithic assembly of tissues, but is instead a true ecosystem composed of human cells, bacteria, fungi, algae, and viruses. As such, humans are not unlike other complex ecosystems containing microbial assemblages observed in the marine and earth environments. They all share a basic functional principle: Chemical communication is the universal language that allows such groups to properly function together. These chemical networks regulate interactions like metabolic exchange, antibiosis and symbiosis, and communication. The National Academies of Sciences, Engineering, and Medicine's Chemical Sciences Roundtable organized a series of four seminars in the autumn of 2016 to explore the current advances, opportunities, and challenges toward unveiling this "chemical dark matter" and its role in the regulation and function of different ecosystems. The first three focused on specific ecosystemsâ€"earth, marine, and humanâ€"and the last on all microbiome systems. This publication summarizes the presentations and discussions from the seminars.
