Tesis sobre el tema "Gene technology"
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Busby, Michele Anne. "Measuring Gene Expression With Next Generation Sequencing Technology". Thesis, Boston College, 2012. http://hdl.handle.net/2345/3145.
Texto completoWhile a PhD student in Dr. Gabor Marth's laboratory, I have had primary responsibility for two projects focused on using RNA-Seq to measure differential gene expression. In the first project we used RNA-Seq to identify differentially expressed genes in four yeast species and I analyzed the findings in terms of the evolution of gene expression. In this experiment, gene expression was measured using two biological replicates of each species of yeast. While we had several interesting biological findings, during the analysis we dealt with several statistical issues that were caused by the experiment's low number of replicates. The cost of sequencing has decreased rapidly since this experiment's design and many of these statistical issues can now practically be avoided by sequencing a greater number of samples. However, there is little guidance in the literature as to how to intelligently design an RNA-Seq experiment in terms of the number of replicates that are required and how deeply each replicate must be sequenced. My second project, therefore, was to develop Scotty, a web-based program that allows users to perform power analysis for RNA-Seq experiments. The yeast project resulted in a highly accessed first author publication in BMC Genomics in 2011. I have structured my dissertation as follows: The first chapter, entitled General Issues in RNA-Seq, is intended to synthesize the themes and issues of RNA-Seq statistical analysis that were common to both papers. In this section, I have discussed the main findings from the two papers as they relate to analyzing RNA-Seq data. Like the Scotty application, this section is designed to be "used" by wet-lab biologists who have a limited background in statistics. While some background in statistics would be required to fully understand the following chapters, the essence of this background can be gained by reading this first chapter. The second and third chapters contain the two papers that resulted from the two RNA-Seq projects. Each chapter contains both the original manuscript and original supplementary methods and data section. Finally, I include brief summaries of my contributions to the two papers on which I was a middle author. The first was a functional analysis of the genomic regions affected by mobile element insertions as a part of Chip Stewart's paper with the 1000 Genome Consortium. This paper was published in Plos Genetics. The second was a cluster analysis of microarray gene expression in Toxoplasma gondii, which was included as part of Alexander Lorestani et al.'s paper, Targeted proteomic dissection of Toxoplasma cytoskeleton sub-compartments using MORN1. This paper is currently under review. The yeast project was a collaborative effort between Jesse Gray, Michael Springer, and Allen Costa at Harvard Medical School, Jeffery Chuang here at Boston College, and members of the Marth lab. Jesse Gray conceived of the project. While I wrote the draft for the manuscript, many people, particularly Gabor Marth, provided substantial guidance on the actual text. I conceived of and implemented Scotty and wrote its manuscript with only editorial assistance from my co-authors. I produced all figures for the two manuscripts. Chip Stewart provided extensive guidance and mentorship to me on all aspects of my statistical analyses for both projects
Thesis (PhD) — Boston College, 2012
Submitted to: Boston College. Graduate School of Arts and Sciences
Discipline: Biology
Heywood, Jacqualine y n/a. "'Talking' and 'doing' gene technology politics: a policy analysis". Griffith University. Australian School of Environmental Studies, 2004. http://www4.gu.edu.au:8080/adt-root/public/adt-QGU20041029.100010.
Texto completoNaujoks, Daniel. "Developing a gene targeting technology for Anopheles gambiae mosquitoes". Thesis, Imperial College London, 2012. http://hdl.handle.net/10044/1/9864.
Texto completoHeywood, Jacqualine. "'Talking' and 'doing' gene technology politics: a policy analysis". Thesis, Griffith University, 2004. http://hdl.handle.net/10072/365762.
Texto completoThesis (PhD Doctorate)
Doctor of Philosophy (PhD)
Australian School of Environmental Studies
Full Text
Fält, Susann. "Analysis of global gene expression in complex biological systems using microarray technology /". Stockholm, 2006. http://diss.kib.ki.se/2006/91-7140-612-3/.
Texto completoHe, Huiqi. "Miniaturized electroporation system for gene transfer using bio-MEMS technology /". View abstract or full-text, 2007. http://library.ust.hk/cgi/db/thesis.pl?BIEN%202007%20HE.
Texto completoWhite, Adam. "Microfluidic technology for high-throughput single cell gene expression analysis". Thesis, University of British Columbia, 2010. http://hdl.handle.net/2429/27838.
Texto completoKotnik, Katarina [Verfasser]. "Gene knockdown in transgenic rats by shRNA technology / Katarina Kotnik". Berlin : Freie Universität Berlin, 2008. http://d-nb.info/1023169665/34.
Texto completoCatteruccia, Flaminia. "Systematic attempts to develop gene transfer technology for anopheline mosquitoes". Thesis, Imperial College London, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.323867.
Texto completoAlete, Julia. "Safe, site-specific gene delivery using ultrasound and microbubble technology". Thesis, Imperial College London, 2008. http://hdl.handle.net/10044/1/4256.
Texto completoRogowski, Wolf. "Key issues in the economic evaluation of gene technology in healthcare /". [S.l. : s.n.], 2007. http://www.gbv.de/dms/zbw/559909535.pdf.
Texto completoVizcaino, Caston Isaac. "Monitoring bacterial physiology during recombinant protein production using reporter gene technology". Thesis, University of Birmingham, 2012. http://etheses.bham.ac.uk//id/eprint/3309/.
Texto completoYoshimura, Shigehiro. "Nano-biology of the Gene : Application of Nano-technology to the Structural and Functional Analysis of the Gene". Kyoto University, 2002. http://hdl.handle.net/2433/149880.
Texto completo0048
新制・課程博士
博士(人間・環境学)
甲第9673号
人博第157号
13||142(吉田南総合図書館)
新制||人||37(附属図書館)
UT51-2002-G431
京都大学大学院人間・環境学研究科人間・環境学専攻
(主査)教授 速水 正憲, 教授 津田 謹輔, 助教授 倉橋 和義, 教授 竹安 邦夫
学位規則第4条第1項該当
McTaggart, Sally. "Retroviral vector production for gene therapy applications". Thesis, University of Birmingham, 2001. http://etheses.bham.ac.uk//id/eprint/7571/.
Texto completoCalus, Szymon Tomasz. "Evaluation of nanopore-based sequencing technology for gene marker based analysis of complex microbial communities : method development for accurate 16S rRNA gene amplicon sequencing". Thesis, University of Glasgow, 2018. http://theses.gla.ac.uk/41086/.
Texto completoThorsén, Jenny, Stina Wahlström, Anna Nilsson, Johanna Öberg y Sebastian Persson. "Immunoassay Applications in Gene and Cell Therapy : A Market Analysis of Companies Conducting Gene and Cell Therapy Product Development". Thesis, Uppsala universitet, Institutionen för biologisk grundutbildning, 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-352164.
Texto completoXie, Dan y 謝丹. "Application of high-throughput tissue microarray technology in cancer research". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2004. http://hub.hku.hk/bib/B30283619.
Texto completoGlare, Eric M. 1965. "The application of competitive PCR technology to asthma research". Monash University, Dept. of Medicine, 2001. http://arrow.monash.edu.au/hdl/1959.1/7975.
Texto completoRuryk, Andriy. "Development of microsystem technology suitable for bacterial identification and gene expression monitoring". [S.l.] : [s.n.], 2005. http://deposit.ddb.de/cgi-bin/dokserv?idn=974502286.
Texto completoHowell, Brandon George. "Gene expression profiling of UV-induced skin cancer using cDNA microarray technology". Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2001. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp05/MQ63108.pdf.
Texto completoJirajaroenrat, Kanya. "Development of ribozyme technology for gene down-regulation in farm animal species". Thesis, University of Nottingham, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.408887.
Texto completoGibbings, J. George. "Transcript profiling in rice using Serial Analysis of Gene Expression (SAGE) technology". Thesis, University of Reading, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.401446.
Texto completoAllum, Nicholas Charles. "Risk, social trust and knowledge : public perceptions of gene technology in Britain". Thesis, London School of Economics and Political Science (University of London), 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.415888.
Texto completoChono, Hideto. "Development of retroviral vector technology and application to HIV-1 gene therapy". Kyoto University, 2012. http://hdl.handle.net/2433/157729.
Texto completoWilliams, Darren William. "Gene transfer and transient expression of transgenes in zebrafish Danio Refrio". Thesis, University of Southampton, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.242103.
Texto completoBelshaw, N. J. "The nuclear matrix and gene expression in Trichoderma reesei". Thesis, University of East Anglia, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.296346.
Texto completoWarnock, James Neill. "Optimisation of retroviral production systems for gene therapy applications". Thesis, University of Birmingham, 2003. http://etheses.bham.ac.uk//id/eprint/3592/.
Texto completoRandel, Melissa. "New Technology Development for Next-Generation Sequencing". Thesis, University of Oregon, 2017. http://hdl.handle.net/1794/22704.
Texto completoCostello, Cait. "Horizontal gene transfer in bacterial co-cultures in micro-fabricated environments". Thesis, University of Birmingham, 2012. http://etheses.bham.ac.uk//id/eprint/3311/.
Texto completoThomas, Alistair Owen. "Detection of bacterial gene expression by a novel isothermic nucleic acid amplification technology". Thesis, University of Bath, 2004. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.410924.
Texto completoZiou, Arisa. "Gene expression of MYPT1 and ROCK1 in endometrial cancer". Thesis, Högskolan i Skövde, Institutionen för hälsovetenskaper, 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:his:diva-19249.
Texto completoChan, V. T. W. "Evaluation of the of the C-HA-RAS in human breast disease by nonisotopic hybridization technology". Thesis, University of Oxford, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.233535.
Texto completoEklöf, Jenny. "Gene technology at stake : Swedish governmental commissions on the border of science and politics". Doctoral thesis, Umeå University, Historical Studies, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-1424.
Texto completoThis thesis examines the Swedish political response to the challenges posed by gene technology, seen through the prism of governmental commissions. It discerns and analyses continuities and changes in the Swedish political conception of gene technology, over the course of two decades, 1980–2000. This is done by thematically following ideas of “risks” and “ethics” as they are represented in the inner workings and reception of three governmental commissions. The Gene-Ethics Commission (1981–1984), the Gene Technology Commission (1990–1992) and the Biotechnology Commission (1997–2000) form the empirical focal points of this analysis. The first two provided preparatory policy proposals that preceded the implementation of the Swedish gene technology laws of 1991 and 1994. The last one aimed at presenting a comprehensive Swedish biotechnology policy for the new millennium.
The study takes into account the role of governmental commissions as arenas where science and politics intersect in Swedish political life, and illuminates how this type of “boundary organisation”, placed on the border of science and politics, impinges on the understanding of the gene technology issue. The commissions have looked into the limits, dangers, possibilities and future applications of gene technology. They have been appointed to deal with the problematic task of distinguishing between what is routine and untested practices, realistic prediction and “science fiction”, what are unique problems and what are problems substantially similar to older ones, what constitutes a responsible approach as opposed to misconduct and what it means to let things “get out of hand” in contrast to being “in control”. Throughout a period of twenty years, media reports have continued to frame the challenges posed by gene technology as a task of balancing risks and benefits, walking the fine line between “frankenfoods” and “miracle drugs”.
One salient problem for the commissions to solve was that science and industry seemed to promote a technology the public opposed and resisted, at least in parts. For both politics and science to gain, or regain, public trust it needed to demonstrate that risks – be it environmental, ethical or health related ones – were under control. Under the surface, it was much more complicated than “science helping politics” to make informed and rational decisions on how to formulate a regulatory policy. Could experts be trusted to participate in policy-making in a neutral way and was it not important, in accordance with democratic norms, to involve the public?
Tong, Lily. "Probing the function of RNase E family using biochemical techniques and gene array technology". Thesis, University of Leeds, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.414514.
Texto completoBerger, Adeline. "Gene therapy for autosomal dominant retinitis pigmentosa : repair of rhodopsin mRNA by SMaRT technology". Thesis, Paris 6, 2014. http://www.theses.fr/2014PA066190/document.
Texto completoRetinitis pigmentosa is an hereditary retinal dystrophy involving degeneration of photoreceptors leading to blindness and for which there is currently no treatment. The most frequent cause of autosomal dominant forms of the disease is a point mutation in the rhodopsin gene (RHO). Therapeutic strategy should both suppress mutant protein expression and restore that of the normal one to physiologic level to prevent photoreceptor degeneration. My PhD project was to repair RHO pre-mRNA by SMaRT (Spliceosome Mediated RNA Trans-splicing) technology. This implies to introduce by gene transfer into the target cell an exogenous RNA, called PTM for Pre-mRNA Trans-splicing Molecule. This one was able to promote a splice reaction in trans, leading to the replacement of the mutated exons. We designed 20 different PTM and obtained in vitro a maximum trans-splicing rate of 40% after transient co-transfection of PTM and RHO constructs in HEK293T cells. We then created WT or mutated RHO stable expression cell lines by lentiviral transduction. Mutation induced retention of the protein into the cytoplasm, while the WT RHO was localized to the plasma membrane. We observed that the PTM transfection in the mutated RHO cell line induced trans-splicing, which was able to partially restore localization to the plasma membrane of repaired RHO. We then tested trans-splicing in vivo in mRho+/- RHO P347S+ mice, a humanized heterozygous mouse model of retinitis pigmentosa. After subretinal injection of AAV2/8-bRho-PTM we observed that trans-splicing occurred in vivo. Unfortunately we did not observe by SD-OCT (a technology that we improve in this project) any rescue of the degenerative phenotype
Aoyama, Teruyoshi. "Improvement of technology for non-viral gene delivery system using DNA-cationized gelatin complex". Kyoto University, 2003. http://hdl.handle.net/2433/148731.
Texto completoEklöf, Jenny. "Gene technology at stake : Swedish governmental commissions on the border of science and politics /". Umeå : Department of Historical Studies, Umeå University, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-1424.
Texto completoMori, Tomoaki. "Application of artificial zinc-finger protein technology: gene regulation and site-specific DNA cleavage". 京都大学 (Kyoto University), 2010. http://hdl.handle.net/2433/120946.
Texto completoRagan, Paula Marie. "The effect of mechanical compression on chondrocyte gene expression". Thesis, Massachusetts Institute of Technology, 1999. http://hdl.handle.net/1721.1/85347.
Texto completoIncludes bibliographical references (leaves 115-122).
by Paula M. Ragan.
Ph.D.
Holland, Chad D. (Chad Darrel). "Personalized medicine, population genetics and privacy : an empirical study of international gene banks". Thesis, Massachusetts Institute of Technology, 2005. http://hdl.handle.net/1721.1/33089.
Texto completoIncludes bibliographical references.
The promise of personalized medicine lies in its potential to fundamentally change healthcare. In the past, pharmaceuticals were prescribed on a "one size fits all" basis-patients with certain disease phenotypes were given what were thought to be appropriate drugs. There is growing evidence however that the effectiveness of these drugs may differ by individual and by sub-group; presumably due to fundamental genetic differences in disease and metabolic pathways. Drugs like Herceptin, Gleevec and Iressa are part of an emerging trend in the biopharmaceutical arena of drugs that are accompanied by genetic diagnostic tests and prescribed only for patients with genotypes in which the agents are most effective.
by Chad D. Holland.
S.M.M.O.T.
S.M.
Bhattacharya, Kanishka. "Gene x gene interactions in genome wide association studies". Thesis, University of Oxford, 2014. http://ora.ox.ac.uk/objects/uuid:6cb7ab29-90df-4d70-bc2f-531f874b79d0.
Texto completoGerber, Georg Kurt 1970. "Computational discovery of gene modules, regulatory networks and expression programs". Thesis, Massachusetts Institute of Technology, 2007. http://hdl.handle.net/1721.1/42201.
Texto completoIncludes bibliographical references (p. 163-181).
High-throughput molecular data are revolutionizing biology by providing massive amounts of information about gene expression and regulation. Such information is applicable both to furthering our understanding of fundamental biology and to developing new diagnostic and treatment approaches for diseases. However, novel mathematical methods are needed for extracting biological knowledge from high-dimensional, complex and noisy data sources. In this thesis, I develop and apply three novel computational approaches for this task. The common theme of these approaches is that they seek to discover meaningful groups of genes, which confer robustness to noise and compress complex information into interpretable models. I first present the GRAM algorithm, which fuses information from genome-wide expression and in vivo transcription factor-DNA binding data to discover regulatory networks of gene modules. I use the GRAM algorithm to discover regulatory networks in Saccharomyces cerevisiae, including rich media, rapamycin, and cell-cycle module networks. I use functional annotation databases, independent biological experiments and DNA-motif information to validate the discovered networks, and to show that they yield new biological insights. Second, I present GeneProgram, a framework based on Hierarchical Dirichlet Processes, which uses large compendia of mammalian expression data to simultaneously organize genes into overlapping programs and tissues into groups to produce maps of expression programs. I demonstrate that GeneProgram outperforms several popular analysis methods, and using mouse and human expression data, show that it automatically constructs a comprehensive, body-wide map of inter-species expression programs.
(cont.) Finally, I present an extension of GeneProgram that models temporal dynamics. I apply the algorithm to a compendium of short time-series gene expression experiments in which human cells were exposed to various infectious agents. I show that discovered expression programs exhibit temporal pattern usage differences corresponding to classes of host cells and infectious agents, and describe several programs that implicate surprising signaling pathways and receptor types in human responses to infection.
by Georg Kurt Gerber.
Ph.D.
Pierce, Lain Xylia. "Analysis of Rhythmic Gene Transcription using the TimeR, a Novel Technology to Capture Zebrafish Embryos". Cleveland, Ohio : Case Western Reserve University, 2008. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=case1212770242.
Texto completoGlover, Hilary Rose. "The design and use of reporter gene technology in the understanding of oestrogen receptor function". Thesis, Queen Mary, University of London, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.417216.
Texto completoMitra, Robi David. "Polony sequencing : DNA sequencing technology and a computational analysis reveals chromosomal domains of gene expression". Thesis, Massachusetts Institute of Technology, 2000. http://hdl.handle.net/1721.1/8797.
Texto completoIncludes bibliographical references.
The first part of this thesis describes the development of polony sequencing, a sequencing technology in which DNA is cloned, amplified and sequenced in a polymer matrix. A complex library of one to ten million linear DNA molecules is amplified by performing polymerase chain reaction (PCR) in a thin polyacrylamide film poured on a glass microscope slide. The polyacrylamide matrix retards the diffusion of the DNA molecules so that each amplification product remains localized near its parent molecule. At the end of the reaction, a number of polymerase colonies, or "polonies", have formed, each one grown from a single template molecule. As many as 5 million clones can be amplified in parallel on a single slide. By including an acrydite modification at the 5' end of one of the PCR primers, the amplified DNA will be covalently attached to the polyacrylamide matrix, allowing further enzymatic manipulations to be performed on all clones simultaneously. Also described in this thesis is my progress in development of a protocol to sequence the polonies by repeated cycles of extension with fluorescent deoxynucleotide. Because polony sequencing is inherently parallel, and sub-picoliter volumes are used for each reaction, the technology should be substantially faster and cheaper than existing methods. Applications for polony sequencing such as gene expression analysis, SNP discovery, and SNP screening will also be discussed. The second part of this thesis describes a computational analysis that tests the hypothesis that chromosomal position affects gene expression. It is shown that, throughout the genome, genes lying close together on the same chromosome often show significant coexpression. This coexpression is independent of the orientation of genes to each other, but is dependent on the distance between genes. In several cases where adjacent genes show highly correlated expression, the promoter of only one of the genes contains an upstream activating sequence (UAS) known to be associated with the expression pattern. These results suggest that in certain regions of the genome a single transcription factor binding site may regulate several genes. It is also shown that evolution may take advantage of this phenomenon by keeping genes with similar functions in adjacent positions along the chromosomes. The techniques that are presented provide a computational method to delineate the locations of chromosomal domains and identify the boundary elements that flank them.
Robi David Mitra.
Ph.D.
Leandersson, Viktor. "GENVO: GENE EVOLUTIONVISUALIZATION : A 3D reconciliation software for phylogenies". Thesis, KTH, Medieteknik och interaktionsdesign, MID, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-189179.
Texto completoAtt förstå geners evolution är relevant för flertalet områden, inte minst för attutforska den mänskliga kroppen. Ända sedan Darwin har forskare visualiseratevolution med hjälp av rotade binära träd. Dock utvecklas gener begränsat till hurdess arter utvecklats, och denna relation är av intresse att förstå och utforska. Attgöra en sammanslagning av ett gen-träd med sitt art-träd (reconciliation) är ett sättatt visualisera relationen, men lösningen blir ofta väldigt rörig och svårförståelig.Därför presenterar jag i detta examensarbete Genvo, ett reconciliation programsom använder en simpel träd-layout-algoritm för att visualisera geners evolution irelation till arters evolution i 3D. Jag presenterar även en gedigen problemkarakteriseringför de uppgifter forskarna utför när de jobbar med dagensvisualiseringsverktyg. En prototyp av Genvo testades sedan i en pairanalytics studie, med en testgrupp bestående av fem par. Studien är sedan jämfördmed en workshop där tre testpersoner, med tidigare erfarenhet, jobbade medsamma uppgifter. Analysen av studiernas resultat tyder på en snabbare förståelseav datan när användarna använde sig av Genvo.
Burkhart, Tandace L. "The Search for Novel Sponge genes: Comparative Analysis of Gene Expression in Multiple Sponges". NSUWorks, 2012. http://nsuworks.nova.edu/occ_stuetd/194.
Texto completoCaldarelli, Antonio. "A study on endocrine disrupters in the environment through the microarray technology". Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2007. http://nbn-resolving.de/urn:nbn:de:swb:14-1175076879253-16448.
Texto completoKing, Kevin R. (Kevin Robert) 1976. "High-throughput microfluidic living cell arrays for spatiotemporal gene expression profiling". Thesis, Massachusetts Institute of Technology, 2008. http://hdl.handle.net/1721.1/43809.
Texto completoPage 146 blank.
Includes bibliographical references.
The cellular microenvironment is remarkably complex. In the small space near each cell, growth factors are liberated from extracellular matrix, cytokines are secreted from neighboring cells, and hormones arrive from distant organs. These spatially and temporally diverse cues are integrated by signal transduction cascades to modulate the activity of transcription factors, the principle regulators of gene expression. To date, experimental investigation of spatial and temporal transcription factor activation patterns has been limited by the use of destructive measurement techniques that require averaging responses over large cell populations. Similarly, control of complex microenvironments has been limited by the use of static tissue culture platforms. This thesis describes development of a high-throughput experimental platform called the microfluidic living cell array (mLCA) that combines fluidically-addressable cell arrays with a library of GFP reporter cells to enable nondestructive spatiotemporal gene expression profiling in living cells. The first section describes construction of the GFP reporter library and the development of methodologies for performing routine seeding and culture of cells in microfluidic channels. Microfluidic circuits are then designed to achieve parallel control of soluble stimulus concentration and timing for delivery to downstream cells. A novel "Flow-encoded Switching" (FES) design strategy is introduced to control simultaneous delivery of temporally distinct stimulus patterns using a single input. These circuits are demonstrated by profiling dynamic transcriptional responses to cytokine stimulation, and in each case, cell responses are found to depend quantitatively and qualitatively on the timing of the stimulus.
(cont.) The second section describes development of a two-dimensional valve-controlled mLCA for simultaneously profiling the entire transcriptional reporter library in response to a panel of stimuli. Integrated microvalve arrays control row-seeding and column-stimulation of 256 nanoliter-scale bioreactors, creating a high density matrix of stimulus-response experiments. The platform is demonstrated in the context of the hepatocyte stress response by collecting -5000 single-time-point measurements in each automated and unattended experiment. Results from these studies revealed a novel relationship between TNF-alpha and heat shock response activation, and more generally, illustrated that a single cytokine can activate multiple transcription factors with distinct dynamics. The third section transitions from temporal to spatial profiling and describes discovery and exploration of a spatially heterogeneous gene expression pattern in the innate immune system. Using a stable monoclonal ISRE-GFP reporter, double-stranded DNA (dsDNA) stimulation is found to result in 'colonylike' patterns of reporter activity in an otherwise confluent monolayer. Cell sorting and expression profiling reveal that activated reporter colonies are functionally distinct from their non-activated neighbors, and that colonies are responsible for the majority of cytokine and chemokine expression, including the potent antiviral interferon-beta. Using a novel transplant co-culture experiment, colonies are shown to form by contact-dependent intercellular communication and furthermore, this communication is found to depend on gap junctions. In summary, this thesis introduces promising new tools for conducting high-throughput investigations of spatiotemporal gene expression patterns in living cells, and it provides evidence for a novel dsDNA-induced intercellular communication mechanism that amplifies innate immune responses.
by Kevin R. King.
Ph.D.
Sakian, Sina. "Investigation of methylation and gene expression in placenta of pregnancies conceived by assisted reproductive technology (ART)". Thesis, University of British Columbia, 2011. http://hdl.handle.net/2429/31191.
Texto completo