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 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 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 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 CRISPR written by Rodolphe Barrangou and published by John Wiley & Sons. This book was released on 2022-09-14 with total page 308 pages. Available in PDF, EPUB and Kindle. Book excerpt: “An excellent compendium of all things CRISPR from some of the leading minds in the field. With thorough coverage from every angle and beautifully detailed illustrations, this book is not to be missed!” Jennifer A. Doudna, Professor of Chemistry, Biochemistry & Molecular Biology, UC Berkeley; Founder, Innovative Genomics Institute; Nobel Laureate and coinventor of CRISPR technology “This journey through CRISPR biology and several of its breakthrough applications offers an exciting glimpse into one of the most beautiful and compelling fields in the life sciences.” David R. Liu, Director of the Merkin Institute at the Broad Institute of MIT and Harvard; Professor of Chemistry and Chemical Biology at Harvard University; coinventor of base editing and prime editing “A must read! The CRISPR topics, written by world-leading experts, span from the fascinating mechanistic underpinnings to the ingenious applications. One can read from start to finish or pick and choose themes. Either way, the book delivers utterly enjoyable learning!” Bonnie Bassler, Squibb Professor and Chair, Princeton University Department of Molecular Biology; Howard Hughes Medical Institute Investigator CRISPR-Cas systems have revolutionized the science of gene editing and their possible applications continue to expand, from basic research to potentially groundbreaking medical and commercial uses. Led by a distinguished team of editors, CRISPR: Biology and Applications explores the subject matter needed to delve into this fascinating area. Topics covered include: Classification and molecular mechanisms of CRISPR-Cas systems CRISPR-Cas evolution, regulation, expression, and function Uses for gene editing and modulation of gene expression CRISPR-based antimicrobials and phage resistance for medical and industrial purposes Written by internationally renowned authors, CRISPR: Biology and Applications serves as both an introductory guide for those new to the field and an authoritative reference for seasoned researchers whose work touches this evolving and headline- making science.
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 Stress and Environmental Regulation of Gene Expression and Adaptation in Bacteria written by Frans J. de Bruijn and published by John Wiley & Sons. This book was released on 2016-07-13 with total page 1472 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bacteria in various habitats are subject to continuously changing environmental conditions, such as nutrient deprivation, heat and cold stress, UV radiation, oxidative stress, dessication, acid stress, nitrosative stress, cell envelope stress, heavy metal exposure, osmotic stress, and others. In order to survive, they have to respond to these conditions by adapting their physiology through sometimes drastic changes in gene expression. In addition they may adapt by changing their morphology, forming biofilms, fruiting bodies or spores, filaments, Viable But Not Culturable (VBNC) cells or moving away from stress compounds via chemotaxis. Changes in gene expression constitute the main component of the bacterial response to stress and environmental changes, and involve a myriad of different mechanisms, including (alternative) sigma factors, bi- or tri-component regulatory systems, small non-coding RNA’s, chaperones, CHRIS-Cas systems, DNA repair, toxin-antitoxin systems, the stringent response, efflux pumps, alarmones, and modulation of the cell envelope or membranes, to name a few. Many regulatory elements are conserved in different bacteria; however there are endless variations on the theme and novel elements of gene regulation in bacteria inhabiting particular environments are constantly being discovered. Especially in (pathogenic) bacteria colonizing the human body a plethora of bacterial responses to innate stresses such as pH, reactive nitrogen and oxygen species and antibiotic stress are being described. An attempt is made to not only cover model systems but give a broad overview of the stress-responsive regulatory systems in a variety of bacteria, including medically important bacteria, where elucidation of certain aspects of these systems could lead to treatment strategies of the pathogens. Many of the regulatory systems being uncovered are specific, but there is also considerable “cross-talk” between different circuits. Stress and Environmental Regulation of Gene Expression and Adaptation in Bacteria is a comprehensive two-volume work bringing together both review and original research articles on key topics in stress and environmental control of gene expression in bacteria. Volume One contains key overview chapters, as well as content on one/two/three component regulatory systems and stress responses, sigma factors and stress responses, small non-coding RNAs and stress responses, toxin-antitoxin systems and stress responses, stringent response to stress, responses to UV irradiation, SOS and double stranded systems repair systems and stress, adaptation to both oxidative and osmotic stress, and desiccation tolerance and drought stress. Volume Two covers heat shock responses, chaperonins and stress, cold shock responses, adaptation to acid stress, nitrosative stress, and envelope stress, as well as iron homeostasis, metal resistance, quorum sensing, chemotaxis and biofilm formation, and viable but not culturable (VBNC) cells. Covering the full breadth of current stress and environmental control of gene expression studies and expanding it towards future advances in the field, these two volumes are a one-stop reference for (non) medical molecular geneticists interested in gene regulation under stress.
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 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 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 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 CRISPR Cas Enzymes written by and published by Academic Press. This book was released on 2019-01-25 with total page 454 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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
Download or read book Gene Expression Systems written by Joseph M. Fernandez and published by Academic Press. This book was released on 1999 with total page 504 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recombinant gene expression is the fastest growing area in the study of molecular biology. By the time the Human Genome Project is completed (~2002), several thousand sequences will be known, but the purpose of the resultant expression products will remain a mystery. Gene discovery requires efficient expression systems for determining the structure and function of gene products. Gene Expression Systems covers a variety of promoters and host organisms that researchers can tailor to their specific needs.
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 Insect Biotechnology written by Andreas Vilcinskas and published by Springer Science & Business Media. This book was released on 2010-10-27 with total page 269 pages. Available in PDF, EPUB and Kindle. Book excerpt: The book provides a fascinating overview about current and sophisticated developments in applied entomology that are powered by molecular biology and that can be summarized under a novel term: insect biotechnology. By analogy with the application of powerful molecular biological tools in medicine (red biotechnology), plant protection (green biotechnology) and industrial processing (white biotechnology), insect biotechnology (yellow biotechnology) provides novel tools and strategies for human welfare and nutrition. Insect Biotechnology has emerged as a prospering discipline with considerable economic potential, and encompasses the use of insect model organisms and insect-derived molecules in medical research as well as in modern plant protection measures.
Download or read book Gene Editing in Plants written by and published by Academic Press. This book was released on 2017-07-14 with total page 266 pages. Available in PDF, EPUB and Kindle. Book excerpt: Gene Editing in Plants, Volume 149 aims to provide the reader with an up-to-date survey of cutting-edge research with gene editing tools and an overview of the implications of this research on the nutritional quality of fruits, vegetables and grains. New chapters in the updated volume include topics relating to Genome Engineering and Agriculture: Opportunities and Challenges, the Use of CRISPR/Cas9 for Crop Improvement in Maize and Soybean, the Use of Zinc-Finger Nucleases for Crop Improvement, Gene Editing in Polyploid Crops: Wheat, Camelina, Canola, Potato, Cotton, Peanut, Sugar Cane, and Citrus, and Gene Editing With TALEN and CRISPR/Cas in Rice. This ongoing serial contain contributions from leading scientists and researchers in the field of gene editing in plants who describe the results of their own research in this rapidly expanding area of science. - Shows the importance of revolutionary gene editing technology on plant biology research and its application to agricultural production - Provides insight into what may lie ahead in this rapidly expanding area of plant research and development - Contains contributions from major leaders in the field of plant gene editing
