Development Of Molecular And Cellular Imaging Tools To Evaluate Gene And Cell Based Therapeutic Strategies In Vivo
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Development Of Molecular And Cellular Imaging Tools To Evaluate Gene And Cell Based Therapeutic Strategies In Vivo
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Author : Jixiang Xia
language : en
Publisher:
Release Date : 2011
Development Of Molecular And Cellular Imaging Tools To Evaluate Gene And Cell Based Therapeutic Strategies In Vivo written by Jixiang Xia and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2011 with Bioluminescence categories.
Molecular imaging modalities are important tools to evaluate the efficacy of gene delivery systems and cell-based therapies. Development and application of these modalities will advance our understanding of the mechanism of transgene expression and cell fate and functions. Physical gene transfer methods hold many advantages over viral vectors among gene therapeutic strategies. Here, we evaluated the efficacy of biolistic ("gene gun") gene targeting to tissues with non-invasive bioluminescence imaging (BLI) methods. Plasmids carrying the firefly luciferase reporter gene were transfected into mouse skin and liver using biolistics, and BLI was measured at various time points after transfer. With optimized DNA loading ratio (DLRs), reporter gene expression reached to peak 1day after transfer to mouse skin, and the maximum depth of tissue penetration was between 200-300[micrometer]. Similar peak expression of reporter gene was found in mouse liver but the expression was relatively stable 4-8 days post-biolistic gene transfer and remained for up to two weeks afterward. Our results demonstrated BLI was an efficient strategy for evaluation of reporter gene expression in the same animals over a period of up to two weeks in vivo. Different tissues showed different expression kinetics, suggesting that this is an important parameter to consider when developing gene therapy strategies for different target tissues. We also employed BLI to measure differentiation of mouse embryonic stem (ES) cells into beating cardiomyocytes in vitro and in vivo. A subset of these cardiomyocytes appears to be derived from an adrenergic lineage that ultimately contribute to substantial numbers of cardiomyocytes primarily on the left side of the heart. At present, it is unclear what the precise role of these cardiac adrenergic cells is with respect to heart development, though it is known that adrenergic hormones (adrenaline and noradrenaline) are essential for embryonic development since mice lacking them die from apparent heart failure during the prenatal period. To identify and characterize cardiac adrenergic cells, we developed a novel mouse genetic model in which the nuclear-localized enhanced green fluorescent protein (nEGFP) reporter gene was targeted to the first exon of the Phenylethanoamine N-transferase (Pnmt) gene, which encodes for the enzyme that converts noradrenaline to adrenaline, and hence serves as a marker for adrenergic cells. Our results demonstrate this knock-in strategy effectively marked adrenergic cells in both fetal and adult mice. Expression of nEGFP was found in Pnmt-positive cells of the adult adrenal medulla, as expected. Pnmt-nEGFP expression also recapitulated endogenous Pnmt expression in the embryonic mouse heart. In addition, nEGFP and Pnmt expression were induced in parallel during differentiation of pluripotent mouse ES cells into beating cardiomyocytes. This new mouse genetic model provides a useful new tool for studying the properties of adrenergic cells in different tissues. We also identified two limitations of the Pnmt-nEGFP model. One is that the amount of nEGFP expressed within individual adrenergic cells was highly variable. Secondly, expression of nEGFP in the embryonic heart was of low abundance and difficult to distinguish from background autofluorescence. To overcome these limitations, we developed two alternative genetic models to investigate adrenergic cells: (1) Mouse embryonic stem cells, which have been previously targeted with Pnmt-Cre recombinase gene, were additionally targeted with a dual reporter plasmid which covered both a loxP-flanked cDNA of red fluorescence protein (HcRed) and also EGFP. Under the undifferentiated status, cells emit red fluorescence as transcription stops before EGFP coding sequence. After differentiation into beating cardiomyoctyes, some cells switch fluorescence from red to green, indicating that excision of loxP-flanked sequences by Cre since Pnmt had been activated. (2) A surface marker, truncated low-affinity nerve growth factor receptor ([delta]LNGFR) was used as the reporter gene as cells expressing this marker can be enriched by magnetic-activated cell sorting (MACS), a potentially efficient way to yield highly purified positive cells at low input abundance in a population. Through a series of subcloning steps, the targeting construct, Pnmt-[delta]LNGFR-Neo-DTA was created and electroporated into 7AC5EYFP embryonic stem cells. Correctly targeted cells were selected by positive and negative screening. These cells provide a new tool with which to identify, isolate, and characterize the function of adrenergic cells in the developing heart, adrenal gland, and other tissues where adrenergic cells make important contributions.
Protein Engineering
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Author : Huimin Zhao
language : en
Publisher: John Wiley & Sons
Release Date : 2021-08-23
Protein Engineering written by Huimin Zhao 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 2021-08-23 with Science categories.
A one-stop reference that reviews protein design strategies to applications in industrial and medical biotechnology Protein Engineering: Tools and Applications is a comprehensive resource that offers a systematic and comprehensive review of the most recent advances in the field, and contains detailed information on the methodologies and strategies behind these approaches. The authors—noted experts on the topic—explore the distinctive advantages and disadvantages of the presented methodologies and strategies in a targeted and focused manner that allows for the adaptation and implementation of the strategies for new applications. The book contains information on the directed evolution, rational design, and semi-rational design of proteins and offers a review of the most recent applications in industrial and medical biotechnology. This important book: Covers technologies and methodologies used in protein engineering Includes the strategies behind the approaches, designed to help with the adaptation and implementation of these strategies for new applications Offers a comprehensive and thorough treatment of protein engineering from primary strategies to applications in industrial and medical biotechnology Presents cutting edge advances in the continuously evolving field of protein engineering Written for students and professionals of bioengineering, biotechnology, biochemistry, Protein Engineering: Tools and Applications offers an essential resource to the design strategies in protein engineering and reviews recent applications.
Molecular Nuclear Medicine
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Author : L.E. Feinendegen
language : en
Publisher: Springer Science & Business Media
Release Date : 2012-12-06
Molecular Nuclear Medicine written by L.E. Feinendegen and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-06 with Medical categories.
Biochemical transparency of the human body is at the doorstep of advanced technology. Toward this goal the book describes relevant isotopic tracer techniques of nuclear medicine. It deals with quantitatively measuring in vivo biochemical reactions as they occur within homeostatic circuits under control by genes and protein interactions. The text indicates how nuclear medicine can aid clinical researchers and practitioners, human geneticists and pharmacologists in understanding (and affecting) gene-phenotype relationships. Experts give background, techniques and examples in an interdisciplinary approach to regional imaging and in vitro analyses of biochemical reactions.
Gene Therapy For Cancer
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Author : Kelly K. Hunt
language : en
Publisher: Springer Science & Business Media
Release Date : 2007-10-26
Gene Therapy For Cancer written by Kelly K. Hunt and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2007-10-26 with Medical categories.
The three sections of this volume present currently available cancer gene therapy techniques. Part I describes the various aspects of gene delivery. In Part II, the contributors discuss strategies and targets for the treatment of cancer. Finally, in Part III, experts discuss the difficulties inherent in bringing gene therapy treatment for cancer to the clinic. This book will prove valuable as the volume of preclinical and clinical data continues to increase.
Molecular Imaging
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Author : Ralph Weissleder
language : en
Publisher: PMPH-USA
Release Date : 2010
Molecular Imaging written by Ralph Weissleder and has been published by PMPH-USA this book supported file pdf, txt, epub, kindle and other format this book has been release on 2010 with Diagnostic imaging categories.
The field of molecular imaging of living subjects have evolved considerably and have seen spectacular advances in chemistry, engineering and biomedical applications. This textbook was designed to fill the need for an authoritative source for this multi-disciplinary field. We have been fortunate to recruit over 80 leading authors contributing 75 individual chapters. Given the multidisciplinary nature of the field, the book is broken into six different sections: "Molecular Imaging technologies", "Chemistry", "Molecular Imaging in Cell and Molecular Biology", "Applications of Molecular Imaging", "Molecular Imaging in Drug Evaluation" with the final section comprised of chapters on computation, bioinformatics and modeling. The organization of this large amount of information is logical and strives to avoid redundancies among chapters. It encourages the use of figures to illustrate concepts and to provide numerous molecular imaging examples.
Molecular Imaging
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Author : Brian D. Ross
language : en
Publisher: Academic Press
Release Date : 2021-08-03
Molecular Imaging written by Brian D. Ross and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021-08-03 with Science categories.
The detection and measurement of the dynamic regulation and interactions of cells and proteins within the living cell are critical to the understanding of cellular biology and pathophysiology. The multidisciplinary field of molecular imaging of living subjects continues to expand with dramatic advances in chemistry, molecular biology, therapeutics, engineering, medical physics and biomedical applications. Molecular Imaging: Principles and Practice, Volumes 1 and 2, Second Edition provides the first point of entry for physicians, scientists, and practitioners. This authoritative reference book provides a comprehensible overview along with in-depth presentation of molecular imaging concepts, technologies and applications making it the foremost source for both established and new investigators, collaborators, students and anyone interested in this exciting and important field. The most authoritative and comprehensive resource available in the molecular-imaging field, written by over 170 of the leading scientists from around the world who have evaluated and summarized the most important methods, principles, technologies and data Concepts illustrated with over 600 color figures and molecular-imaging examples Chapters/topics include, artificial intelligence and machine learning, use of online social media, virtual and augmented reality, optogenetics, FDA regulatory process of imaging agents and devices, emerging instrumentation, MR elastography, MR fingerprinting, operational radiation safety, multiscale imaging and uses in drug development This edition is packed with innovative science, including theranostics, light sheet fluorescence microscopy, (LSFM), mass spectrometry imaging, combining in vitro and in vivo diagnostics, Raman imaging, along with molecular and functional imaging applications Valuable applications of molecular imaging in pediatrics, oncology, autoimmune, cardiovascular and CNS diseases are also presented This resource helps integrate diverse multidisciplinary concepts associated with molecular imaging to provide readers with an improved understanding of current and future applications
Molecular Imaging With Reporter Genes
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Author : Sanjiv Sam Gambhir
language : en
Publisher: Cambridge University Press
Release Date : 2010-05-31
Molecular Imaging With Reporter Genes written by Sanjiv Sam Gambhir and has been published by Cambridge University Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2010-05-31 with Science categories.
Reporter genes have been used for several decades to study regulation of gene expression in vivo. However, it was little more than a decade ago that a new class of reporter genes was developed for imaging molecular events within living subjects. By following the interactions of protein molecules, researchers can resolve the complex chemical pathways that living cells utilise. This book focuses on this group of imaging reporter genes, starting with detailed descriptions of all reporter genes from different imaging modalities, including optical, MRI, and radionuclide-based imaging. Key scientists in the field explain how to enhance reporter gene imaging utility through instrumentation and the various applications of this technology. This is the first comprehensive book on all aspects of reporter gene imaging, detailing what is known in the field and future goals for research. Investigators in biomedical sciences, physicians, and the biotechnology and pharmaceutical industries will benefit from topics covered.
Nanoscale Technology In Biological Systems
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Author : Ralph S. Greco
language : en
Publisher: CRC Press
Release Date : 2004-12-20
Nanoscale Technology In Biological Systems written by Ralph S. Greco and has been published by CRC Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2004-12-20 with Medical categories.
Nanoscale Technology in Biological Systems reviews recent accomplishments in the field of nanobiology and introduces the application of nanoscale matrices to human biology. It focuses on the applications of nanotechnology fabrication to biomedical devices and discusses new physical methods for cell isolation and manipulation and intracellular commu
Molecular Biology Of The Cell
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Author : Bruce Alberts
language : en
Publisher:
Release Date : 2002
Molecular Biology Of The Cell written by Bruce Alberts and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2002 with Cytology categories.
Cardiovascular Molecular Imaging
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Author : Robert J. Gropler
language : en
Publisher: CRC Press
Release Date : 2007-07-23
Cardiovascular Molecular Imaging written by Robert J. Gropler and has been published by CRC Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2007-07-23 with Medical categories.
Cardiovascular Molecular Imaging is based on a groundbreaking NIH symposium sponsored by the American Society of Nuclear Cardiology. The first all-inclusive guide to the targeted molecular imaging of the cardiovascular system, it includes color illustrations throughout and is packaged with a user-friendly CD-ROM with supplemental material.This refe