Download or read book The Development of Reporter Genes for in Vivo Imaging written by Peter Stephen Patrick and published by . This book was released on 2014 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Development of Reporter Genes for in Vivo Imaging of Staphylococcus Aureus written by Rochelle Marie Lamb and published by . This book was released on 2010 with total page 290 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Molecular Imaging with Reporter Genes written by Sanjiv Sam Gambhir and published by Cambridge University Press. This book was released on 2010-05-31 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Download or read book Reporter Gene Assays written by Robert Damoiseaux and published by Humana Press. This book was released on 2018-04-19 with total page 234 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume explores the applications of reporter gene technology and the methodologies needed for their effective implementation. The chapters in this book cover practical topics such as how to integrate reporter constructs into cellular models, viral delivery, splicing applications, in vivo imaging, and a guide to the use of multi-cistronic constructs. Additionally, chapters also include detailed mechanistic uses of reporter genes in cellular pathways, and a look at project and data management of screening applications. 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. Cutting-edge and thorough, Reporter Gene Assays: Methods and Protocols is a valuable resource for anyone who is interested in learning more about reporter genes.

Download or read book Reporter Genes written by Donald S. Anson and published by Springer Science & Business Media. This book was released on 2007-09-13 with total page 165 pages. Available in PDF, EPUB and Kindle. Book excerpt: Reporter genes have played, and continue to play, a vital role in many areas of biological research by providing a ready means for qualitative and quantitative assessment of the activity of genes and location of gene products in different environments. In Reporter Genes: A Practical Guide, renowned researchers describe practical protocols for experimentation with the most useful reporter genes for mammalian systems that are available, concentrating on those marker genes that are currently most commonly used. Among the topics covered in this volume are methodologies for staining and visualization of ß-galactosidase in embryos and tissues, immunohistochemical detection of ß-galactosidase, detection of reporter gene expression in murine airways, and three dimensional analysis of molecular signals with episcopic imaging techniques. Also covered in this groundbreaking text are fluorescent proteins, detection of GFP during nervous system development, fluorescent protein reporter systems for single cell measurements, and non-invasive imaging of molecular events. Comprehensive and illuminating, Reporter Genes: A Practical Guide will be an oft-used reference for geneticists, molecular engineers, and molecular biologists.

Download or read book Development of Molecular and Cellular Imaging Tools to Evaluate Gene and Cell Based Therapeutic Strategies in Vivo written by Jixiang Xia and published by . This book was released on 2011 with total page 130 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Download or read book Textbook of in vivo Imaging in Vertebrates written by Vasilis Ntziachristos and published by John Wiley & Sons. This book was released on 2007-08-20 with total page 388 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book describes the new imaging techniques being developed to monitor physiological, cellular and subcellular function within living animals. This exciting field of imaging science brings together physics, chemistry, engineering, biology and medicine to yield powerful and versatile imaging approaches. By combining advanced non-invasive imaging technologies with new mechanisms for visualizing biochemical events and protein and gene function, non-invasive vertebrate imaging enables the in vivo study of biology and offers rapid routes from basic discovery to drug development and clinical application. Combined with the availability of an increasing number of animal models of human disease, and the ability to perform longitudinal studies of disease evolution and of the long-term effects of therapeutic procedures, this new technology offers the next generation of tools for biomedical research. Well illustrated, largely in colour, the book reviews the most common and technologically advanced methods for vertebrate imaging, presented in a clear, comprehensive format. The basic principles are described, followed by several examples of the use of imaging in the study of living multicellular organisms, concentrating on small animal models of human diseases. The book illustrates: The types of information that can be obtained with modern in vivo imaging; The substitution of imaging methods for more destructive histological techniques; The advantages conferred by in vivo imaging in building a more accurate picture of the response of tissues to stimuli over time while significantly reducing the number of animals required for such studies. Part 1 describes current techniques in in vivo imaging, providing specialists and laboratory scientists from all disciplines with clear and helpful information regarding the tools available for their specific research field. Part 2 looks in more detail at imaging organ development and function, covering the brain, heart, lung and others. Part 3 describes the use of imaging to monitor various new types of therapy, following the reaction in an individual organism over time, e.g. after gene or cell therapy. Most chapters are written by teams of physicists and biologists, giving a balanced coherent description of each technique and its potential applications.

Download or read book Molecular Diagnostics Promises and Possibilities written by Mousumi Debnath and published by Springer Science & Business Media. This book was released on 2010-01-29 with total page 527 pages. Available in PDF, EPUB and Kindle. Book excerpt: A rapid development in diverse areas of molecular biology and genetic engineering resulted in emergence of variety of tools. These tools are not only applicable to basic researches being carried out world over, but also exploited for precise detection of abnormal conditions in plants, animals and human body. Although a basic researcher is well versed with few techniques used by him/her in the laboratory, they may not be well acquainted with methodologies, which can be used to work out some of their own research problems. The picture is more blurred when the molecular diagnostic tools are to be used by physicians, scientists and technicians working in diagnostic laboratories in hospitals, industry and academic institutions. Since many of them are not trained in basics of these methods, they come across several gray areas in understanding of these tools. The accurate application of molecular diagnostic tools demands in depth understanding of the methodology for precise detection of the abnormal condition of living body. To meet the requirements of a good book on molecular diagnostics of students, physicians, scientists working in agricultural, veterinary, medical and pharmaceutical sciences, it needs to expose the reader lucidly to: Give basic science behind commonly used tools in diagnostics Expose the readers to detailed applications of these tools and Make them aware the availability of such diagnostic tools The book will attract additional audience of pathologists, medical microbiologists, pharmaceutical sciences, agricultural scientists and veterinary doctors if the following topics are incorporated at appropriate places in Unit II or separately as a part of Unit-III in the book. Molecular diagnosis of diseases in agricultural crops Molecular diagnosis of veterinary diseases. Molecular epidemiology, which helps to differentiate various epidemic strains and sources of disease outbreaks. Even in different units of the same hospital, the infections could be by different strains of the same species and the information becomes valuable for infection control strategies. Drug resistance is a growing problem for bacterial, fungal and parasitic microbes and the molecular biology tools can help to detect the drug resistance genes without the cultivation and in vitro sensitivity testing. Molecular diagnostics offers faster help in the selection of the proper antibiotic for the treatment of tuberculosis, which is a major problem of the in the developing world. The conventional culture and drug sensitivity testing of tuberculosis bacilli is laborious and time consuming, whereas molecular diagnosis offers rapid drug resistant gene detection even from direct clinical samples. The same approach for HIV, malaria and many more diseases needs to be considered. Molecular diagnostics in the detection of diseases during foetal life is an upcoming area in the foetal medicine in case of genetic abnormalities and infectious like TORCH complex etc. The book will be equally useful to students, scientists and professionals working in the field of molecular diagnostics.

Download or read book In Vivo Cellular and Molecular Imaging written by Eric T. Ahrens and published by Gulf Professional Publishing. This book was released on 2005-11-09 with total page 310 pages. Available in PDF, EPUB and Kindle. Book excerpt: In recent times there has been an explosive expansion of new imaging methodologies that are capable of visualizing specific populations of cells and molecular events in vivo. Vital imaging enhances our ability to study animal models of human development and disease, such as cancers, cardiovascular disease, diabetes, and Alzheimer's. Furthermore, non-invasive imaging may ultimately be useful for monitoring new generations of clinical molecular and cellular therapeutics, such as those utilizing viral vectors and stem cells. These new capabilities have been facilitated by the development of new imaging probes or reagents that target specific cell types, are chemically responsive to physiology, or are responsive to the presence of specific molecules, such as nucleic acids or enzymes. This volume provides an introduction to some of the most exciting methods and applications of emerging non-invasive imaging technologies using magnetic resonance imaging (MRI), positron emission tomography (PET), and various biophotonic approaches. Highlighted, are recent developments in reagent design that impart unique abilities to these imaging modalities to elucidate biological processes in vivo. * Includes 9 chapters by expert researchers in the field of imaging * Introduces new methods and applications of non-invasive imaging technologies * Covers ermerging topics in imaging such as in vivo cell cancer cells, imaging of autoimmune diseases, and maganetic resonance imaging

Download or read book Encyclopedia of Cancer written by Manfred Schwab and published by Springer Science & Business Media. This book was released on 2008-09-23 with total page 3307 pages. Available in PDF, EPUB and Kindle. Book excerpt: This comprehensive encyclopedic reference provides rapid access to focused information on topics of cancer research for clinicians, research scientists and advanced students. Given the overwhelming success of the first edition, which appeared in 2001, and fast development in the different fields of cancer research, it has been decided to publish a second fully revised and expanded edition. With an A-Z format of over 7,000 entries, more than 1,000 contributing authors provide a complete reference to cancer. The merging of different basic and clinical scientific disciplines towards the common goal of fighting cancer makes such a comprehensive reference source all the more timely.

Download or read book From Morphological Imaging to Molecular Targeting written by Markus Schwaiger and published by Springer Science & Business Media. This book was released on 2013-03-09 with total page 218 pages. Available in PDF, EPUB and Kindle. Book excerpt: The current progress in molecular medicine allows the identification of a plethora of new and often human-specific drug targets. An early in vivo validation of specific ligands binding to these targets in humans is needed to assess their potential for targeted imaging and radiotherapy. Radiopharmaceuticals are uniquely suitable for such target validation studies. The purpose of the Ernst Schering Research Foundation Workshop 48 was to offer a forum for an open exchange on the state of the art in the early development of such radiopharmaceuticals. Experts from academia, industry and regulatory authorities provided contributions covering the identification of targets, the necessary preclinical studies on the safety of ligands as well as their validation in human clinical trials.

Download or read book Bioluminescence and Chemiluminescence written by Marlene A. Deluca and published by Elsevier. This book was released on 2013-09-11 with total page 813 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioluminescence and Chemiluminescence: Basic Chemistry and Analytical Applications is a compendium of papers presented at the second International Symposium on Analytical Applications of Bioluminescene and Chemiluminescence in San Diego, California on August 26-28, 1980. Part I deals with chemilunescence and excited states including topics on the spectroscopy of the solvent cage — generation and characteristics of the excited state and the three features of chemi- and bioluminescence. Part II deals with events prior to producing luminol-dependent chemiluminescence; this part also examines the effects of stimulants on membrane potential. Part III discusses bacterial bioluminescence and analyzes the properties of a lumazine protein from a bioluminescent bacterium. This part also analyzes accessory enzymes responsible for such bacterial bioluminescence. Part IV examines the chemistry of firefly bioluminescence and presents the formula of three reactions catalyzed by firefly luciferase. Part V analyzes bioluminescence from other sources such as the earthworm and land snails. Part VI discusses the applications of bioluminescence in clinical chemistry, soil science, and marine biology. Part VII deals with the future uses of bioluminescence, while Part VIII is an abstract of other papers dealing with this subject. This collection can be helpful for biologists, zoologists, micro-biologists, marine biologists, and researchers dealing with bio-chemistry and related disciplines.

Download or read book Molecular Imaging of Cardiac Transgene Expression and Cell Transplantation in Living Subjects written by Joseph C. Wu and published by . This book was released on 2004 with total page 202 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Root Development written by Daniela Ristova and published by Humana Press. This book was released on 2018-03-11 with total page 324 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed volume provides diverse elegant methods, complemented with existing protocols, which are optimized for the current needs in plant root biology as well as for use in plant species other than Arabidopsis thaliana. The collection covers methods ranging from genetic screens, phenotypic analysis, and cell biology methods to systems biology tools and genome-wide approaches. The collection contains a range of complexity from fundamental methods for quantification of different root developmental processes to complex methods that require sophisticated equipment. Written for the highly successful Methods in Molecular Biology series, 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. Authoritative and practical, Root Development: Methods and Protocols serves as a valuable tool for plant biologists specialized in root development, from beginners to experienced researchers in the field.

Download or read book Test No 488 Transgenic Rodent Somatic and Germ Cell Gene Mutation Assays written by OECD and published by OECD Publishing. This book was released on 2011-07-28 with total page 16 pages. Available in PDF, EPUB and Kindle. Book excerpt: This Test Guideline describes an in vivo assay that detects chemicals that may induce gene mutations. In this assay, transgenic rats or mice that contain multiple copies of chromosomally integrated plasmid or phage shuttle vectors are used. The ...

Download or read book Development of Novel Dual mode Radionuclide fluorescence Imaging Platforms for MicroRNA and Hypoxia Imaging written by Eleni Skourti and published by . This book was released on 2018 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This study focuses on imaging specific alterations that are related to cancer progression and more specifically to metastasis. Given the key role that both miRNAs and hypoxia have been found to play in the enhancement of the metastatic potential of cancer cells, the two main goals of this study have been the development of (a) a system to image miRNAs with an implication in metastasis in whole body preclinical models and (b) a new permanent hypoxia switch, which would enable tracking longitudinally cells that have been exposed to hypoxic conditions. Herein, the generation of the miRNA sensor through genetic engineering of plasmid DNA constructs is shown. The sensor involves a human sodium iodide symporter (hIS) based system where hNIS is fused to the green fluorescent protein (GFP) (hNIS-GFP). Additionally, a transcriptional repressor is used to regulate the expression of the reporter. The repressor is modified in a way that renders it sensitive to the presence of the miRNAs of interest. The generation of the miRNA sensor is followed by the confirmation of the functionality of the system. The application of the miRNA sensor is shown, with a focus on a specific triple negative breast cancer cell line, MDA-MB-231. Once the system is shown functional in vitro, it is validated in vivo. Ex vivo histological analysis confirms the in vivo observed functionality of the system. Additionally, an alternative system based on a protease (TEV protease) approach is presented: The generation of the DNA constructs for this approach and the characterisation of the system generated is included hereby. The last part of this study involves the generation of the hypoxia permanent switch. This system is based on a hypoxia sensitive Cre recombinase: when a hypoxic event takes place, Cre is expressed and leads to a recombination of the lox sites surrounding a gene coding for the dsRed protein. This results in a hIS-GP fusion reporter gene coming in frame and being expressed, thus enabling tracking of the cells that have undergone hypoxia. The switch is assessed in a transiently transfected system and then used for evaluation in a stable setup in a human triple negative breast cancer cell line (MDA-MB-231). Finally, the hypoxia permanent switch is characterised in an in vivo system.

Download or read book Molecular Imaging Basic Principles And Applications In Biomedical Research written by Markus Rudin and published by World Scientific. This book was released on 2005-09-29 with total page 564 pages. Available in PDF, EPUB and Kindle. Book excerpt: Molecular imaging is a rapidly emerging field that translates many concepts developed for molecular biology and cellular imaging to the in vivo imaging of intact organisms. The technique allows the study of molecular biological events in their full context and will therefore become an indispensable tool for biomedical research and drug discovery and development.This volume familiarizes the reader with the concepts of imaging and molecular imaging in particular. Basic principles of imaging technologies, reporter moieties for the various imaging modalities and the design of target reporter constructs are described in the first part. The second part illustrates how these tools can be used to visualize relevant molecular events: the biodistribution of drugs/ligands, the expression of drug targets (receptors, enzymes), and the consequences of the molecular drug-target interactions (pathway activations, system responses). A final chapter deals with visualization of cell migration (cell therapies).