Rna Sensing And Programming Platforms For Mammalian Synthetic Biology
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Rna Sensing And Programming Platforms For Mammalian Synthetic Biology
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Author : Breanna Elizabeth DiAndreth
language : en
Publisher:
Release Date : 2019
Rna Sensing And Programming Platforms For Mammalian Synthetic Biology written by Breanna Elizabeth DiAndreth and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019 with categories.
The field of synthetic biology aims to control cellular behavior using programmable gene circuits. Generally these gene circuits sense molecular biomarkers, process these inputs and execute a desired calculated response. This is especially relevant for gene and cell therapies where integrating multiple disease-related inputs and/or sophisticated control could lead to safer and more effective approaches. While mammalian synthetic biology has made great progress, few gene circuit-based therapies have entered the clinic. Regulatory issues aside, this lag may be due to several technical impediments. First, the computing part of circuits is often accomplished via transcriptional regulation, which presents challenges as we move toward the clinic. Second, the field relies on a limited set of sensors; the detection of other types of disease biomarkers will help robustly identify cell state. Finally, the design cycle currently used to develop gene circuits is laborious and slow, which is not suitable for clinical development, especially applications in personalized medicine. In this thesis I describe how I address these three limitations. I develop a new posttranscriptional regulation platform based on RNA cleavage that I term "PERSIST" (Programmable Endonucleolytic RNA Scission-Induced Stability Tuning). CRISPR-specific endonucleases are adapted as RNA-level regulators for the platform and we demonstrate several genetic devices including cascades, feedback, logic functions and a bistable switch. I explore sensor designs for relevant biomolecules including mRNAs, miRNAs and proteins via the PERSIST and other platforms. Finally, I present a "poly-transfection" method, associated advanced data analysis pipelines, and computational models that make circuit engineering faster and more predictive. Taken together, the expanded RNA toolkit that the PERSIST platform offers as well as advancements in sensing and circuit design will enable the more straightforward creation of robust gene circuits for gene and cell therapies.
Synthetic Dna And Rna Programming
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Author : Patrick O'Donoghue
language : en
Publisher:
Release Date : 2019
Synthetic Dna And Rna Programming written by Patrick O'Donoghue and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019 with Biology (General) categories.
Dear Colleagues, Synthetic biology is a broad and emerging discipline that capitalizes on recent advances in molecular biology, genetics, protein and RNA engineering and omics technologies. These technologies have transformed our ability to reveal the biology of the cell and the molecular basis of disease.
Scalable Genetic System Design Using Synthetic Rna Regulators
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Author : Lei Qi
language : en
Publisher:
Release Date : 2012
Scalable Genetic System Design Using Synthetic Rna Regulators written by Lei Qi and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012 with categories.
Our ability to efficiently and predictably program cells is central to the fields of bioengineering and synthetic biology. Once thought to be a passive carrier of genetic information, RNA is now more appreciated as the main organizer of cellular networks. To harness the unique abilities of RNA molecules for programming cells, we show here how to rationally design novel synthetic RNA elements to recapitulate the functions of natural noncoding RNAs (ncRNAs), and how to assemble these synthetic elements into higher-order biological systems. To create synthetic RNA elements, we start with two primary types of ncRNA-mediated natural systems. Both modulate RNA-level regulatory signals encoded in the 5' untranslated region, and are mediated by ncRNAs. In the first system the ncRNA represses transcription elongation, whereas in the second system the ncRNA inhibits translation initiation. To create orthogonal RNA elements that work independently in the same cell, we systematically modify the RNA-RNA interaction in the natural systems. Our characterization results in families of orthogonally acting RNA elements for both transcription and translation controls. Furthermore, we develop mathematical thermodynamic models to predict new RNA elements in silico for translation controls. To engineer synthetic RNAs to sense and integrate cellular signals, we design allosteric RNA chimera molecules by fusing ncRNAs to RNA aptamers. We demonstrate the design principles for creating such chimeric RNA molecules that can sense proteins or small molecules and control transcription or translation. We show that the design strategy is modular, which allows us to reconfigure different ncRNA mutants and RNA aptamers to engineer orthogonal RNA chimeras that respond to different ligands and regulate different gene targets. We further show that multiple RNA chimeras allow logical integration of molecular signals in the same cell for cellular information processing. We assemble multiple synthetic RNA elements to create basic regulatory network motifs. These include independent control, logic control, and cascading control. We characterize the performance and properties of these engineered RNA circuits such as their time response, signal sensitivity, and noises across cell populations. We further explore a strategy that can effectively convert orthogonal translational regulators into orthogonal transcriptional regulators, which can be used to perform multi-input logic computation. In an effort to engineer feedback circuits, we demonstrate the use of translational repressor and activator based on RNA-binding proteins. The designed positive or negative feedback circuits form a basis for programming complex functions. To improve the predictability of engineered biological systems, we develop a synthetic RNA processing platform from the bacterial CRISPR genetic immune pathway. The synthetic RNA processing system can efficiently and specifically cleave desired precursor mRNAs at designed loci. Using this system, we show that transcript cleavage enables quantitative programming of gene expression by modular assembly of promoters, ribosome binding sites, cis regulatory elements, and riboregulators. These basic components can be grouped into multi-gene synthetic operons that behave predictably only after RNA processing. Physical separation of otherwise linked elements within biological assemblies allows design of sophisticated RNA-level regulatory systems that are not possible without it. Thus, our results exemplify a crucial design principle based on controllable RNA processing for improving the modularity and reliability of genetic systems. To sum, our work established bacterial ncRNAs as an intriguing engineering substrate for scalable genetic circuit design and for programming cells. We provide a set of engineering principles for designing synthetic RNA elements as well as using them to sense signals and form genetic circuits. Our RNA-based engineering platform provides a versatile and powerful strategy for designing higher-order cellular information processing and computation systems, which can be readily applied to practical applications including chemical production, environment remediation, and therapeutics.
High Throughput Strategies For The Scalable Generation Of Rna Component Functions
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Author : Joe Chih Yao Liang
language : en
Publisher:
Release Date : 2012
High Throughput Strategies For The Scalable Generation Of Rna Component Functions written by Joe Chih Yao Liang and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012 with Bioengineering categories.
Engineered biological systems hold great promise in providing solutions to many global challenges, including environmental remediation, sustainability, scalable manufacturing, and health and medicine. Synthetic biology is an emerging research field with a primary goal of making the engineering of biology more streamlined and reliable. Recent advances in synthetic RNA biology have led to design of RNA-based gene-regulatory devices from assembly of functional RNA components that encode more basic functions, including sensing, information transmission, and actuation functions. These synthetic RNA control devices allow access and control information on cellular state, thereby advancing our ability to interact with and program biology. A modular ribozyme device platform was recently described to link an aptamer (sensor) to a hammerhead ribozyme (actuator) through a distinct sequence (information transmitter) capable of a strand-displacement event. The utilization of ribozyme as the actuator in the platform, whose mechanism of action is independent of cell-specific machinery, allows transport of the resultant devices to in vitro or different cellular environments. The broad implementation of these devices requires enabling technologies to support efficient generation of new functional RNA components and quantitative tailoring of device regulatory performance for specific cellular applications. Current component generation and device tailoring strategies are limited in their throughputs and efficiencies, and thus have hampered our ability to generate new ribozyme devices for cellular engineering applications. To support scalable generation and tailoring of ribozyme devices, we have described high-throughput in vitro selection and in vivo screening strategies based on the modular ribozyme device platform. We proposed a high-throughput solution-based in vitro selection strategy to generate new sensing functions within the device platform. A high-throughput and quantitative two-color FACS-based screening strategy was developed to complement the in vitro selection strategy by allowing efficient tailoring of device regulatory activities in the cellular environments. We further developed quantitative assays based on the surface plasmon resonance (SPR) technology to allow rapid measurements of the device and component activities. Together, these enabling strategies will offer a scalable and integrated process for the construction and programming of RNA control devices for broad cellular engineering applications, thus laying an important foundation for engineering more complex biological systems.
The Science And Applications Of Synthetic And Systems Biology
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Author : Institute of Medicine
language : en
Publisher: National Academies Press
Release Date : 2011-12-30
The Science And Applications Of Synthetic And Systems Biology written by Institute of Medicine and has been published by National Academies Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2011-12-30 with Science categories.
Many potential applications of synthetic and systems biology are relevant to the challenges associated with the detection, surveillance, and responses to emerging and re-emerging infectious diseases. On March 14 and 15, 2011, the Institute of Medicine's (IOM's) Forum on Microbial Threats convened a public workshop in Washington, DC, to explore the current state of the science of synthetic biology, including its dependency on systems biology; discussed the different approaches that scientists are taking to engineer, or reengineer, biological systems; and discussed how the tools and approaches of synthetic and systems biology were being applied to mitigate the risks associated with emerging infectious diseases. The Science and Applications of Synthetic and Systems Biology is organized into sections as a topic-by-topic distillation of the presentations and discussions that took place at the workshop. Its purpose is to present information from relevant experience, to delineate a range of pivotal issues and their respective challenges, and to offer differing perspectives on the topic as discussed and described by the workshop participants. This report also includes a collection of individually authored papers and commentary.
Synthetic Biology 2 Volumes
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Author : Robert A. Meyers
language : en
Publisher: John Wiley & Sons
Release Date : 2015-10-19
Synthetic Biology 2 Volumes written by Robert A. Meyers and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 2015-10-19 with Science categories.
These two volumes contain a selection of updated articles from the acclaimed Meyers Encyclopedia of Molecular Cell Biology and Molecular Medicine, the most authoritative resource in cell and molecular biology, combined with new articles by "founding fathers" in the field. The work is divided into six sections: + Biological Basis + Modeling + Modular Parts and Circuits + Synthetic Genomes + Diseases and Therapeutics + Chemicals Production. Ideally suited as advanced reading for students and postdocs, and with all current research trends covered by an impressive number of leading figures in the field, this is the first choice reference for research institutions.
Mammalian Synthetic Biology
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Author : Jamie Davies
language : en
Publisher:
Release Date : 2019-12-12
Mammalian Synthetic Biology written by Jamie Davies and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019-12-12 with categories.
Written primarily for mid-to-upper level undergraduates, this primer will introduce students to topics at the forefront of the subject that are being applied to probe biological problems, or to address the most pressing issues facing society. These topics will include those that form thecornerstone of contemporary research, helping students to make the transition to active researcher.This primer introduces the challenges and opportunities of applying synthetic biological techniques to mammalian cells, tissues, and organisms. It covers the special features that make engineering mammalian systems different from engineering bacteria, fungi, and plants, and provides an overview ofcurrent techniques. A variety of cutting-edge examples illustrate the different purposes of mammalian synthetic biology, including pure biomedical research, drug production, tissue engineering, and regenerative medicine.
Cell Free Synthetic Biology
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Author : Jian Li
language : en
Publisher: Frontiers Media SA
Release Date : 2022-01-13
Cell Free Synthetic Biology written by Jian Li and has been published by Frontiers Media SA this book supported file pdf, txt, epub, kindle and other format this book has been release on 2022-01-13 with Science categories.
New Frontiers And Applications Of Synthetic Biology
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Author : Vijai Singh
language : en
Publisher: Academic Press
Release Date : 2022-01-12
New Frontiers And Applications Of Synthetic Biology written by Vijai Singh and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2022-01-12 with Science categories.
New Frontiers and Applications of Synthetic Biology presents a collection of chapters from eminent synthetic biologists across the globe who have established experience and expertise working with synthetic biology. This book offers several important areas of synthetic biology which allow us to read and understand easily. It covers the introduction of synthetic biology and design of promoter, new DNA synthesis and sequencing technology, genome assembly, minimal cells, small synthetic RNA, directed evolution, protein engineering, computational tools, de novo synthesis, phage engineering, a sensor for microorganisms, next-generation diagnostic tools, CRISPR-Cas systems, and more. This book is a good source for not only researchers in designing synthetic biology, but also for researchers, students, synthetic biologists, metabolic engineers, genome engineers, clinicians, industrialists, stakeholders and policymakers interested in harnessing the potential of synthetic biology in many areas. Offers basic understanding and knowledge in several aspects of synthetic biology Covers state-of-the-art tools and technologies of synthetic biology, including promoter design, DNA synthesis, DNA sequencing, genome design, directed evolution, protein engineering, computational tools, phage design, CRISPR-Cas systems, and more Discusses the applications of synthetic biology for smart drugs, vaccines, therapeutics, drug discovery, self-assembled materials, cell free systems, microfluidics, and more
Synthetic Biology
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Author : Huimin Zhao
language : en
Publisher: Academic Press
Release Date : 2013-03-21
Synthetic Biology written by Huimin Zhao and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2013-03-21 with Science categories.
Synthetic Biology provides a framework to examine key enabling components in the emerging area of synthetic biology. Chapters contributed by leaders in the field address tools and methodologies developed for engineering biological systems at many levels, including molecular, pathway, network, whole cell, and multi-cell levels. The book highlights exciting practical applications of synthetic biology such as microbial production of biofuels and drugs, artificial cells, synthetic viruses, and artificial photosynthesis. The roles of computers and computational design are discussed, as well as future prospects in the field, including cell-free synthetic biology and engineering synthetic ecosystems. Synthetic biology is the design and construction of new biological entities, such as enzymes, genetic circuits, and cells, or the redesign of existing biological systems. It builds on the advances in molecular, cell, and systems biology and seeks to transform biology in the same way that synthesis transformed chemistry and integrated circuit design transformed computing. The element that distinguishes synthetic biology from traditional molecular and cellular biology is the focus on the design and construction of core components that can be modeled, understood, and tuned to meet specific performance criteria and the assembly of these smaller parts and devices into larger integrated systems that solve specific biotechnology problems. Includes contributions from leaders in the field presents examples of ambitious synthetic biology efforts including creation of artificial cells from scratch, cell-free synthesis of chemicals, fuels, and proteins, engineering of artificial photosynthesis for biofuels production, and creation of unnatural living organisms Describes the latest state-of-the-art tools developed for low-cost synthesis of ever-increasing sizes of DNA and efficient modification of proteins, pathways, and genomes Highlights key technologies for analyzing biological systems at the genomic, proteomic, and metabolomic levels which are especially valuable in pathway, whole cell, and multi-cell applications Details mathematical modeling tools and computational tools which can dramatically increase the speed of the design process as well as reduce the cost of development