WFRF:(Weibrecht Irene)
Sökning: WFRF:(Weibrecht Irene) > (2011) > Visualizing Interac...
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- Weibrecht, Irene,1983-Uppsala universitet,Molekylära verktyg,Ulf Landegren (författare)
Visualizing Interacting Biomolecules In Situ
- BokEngelska2011
Förlag, utgivningsår, omfång ...
- Uppsala :Acta Universitatis Upsaliensis,2011
- 55 s.
- electronicrdacarrier
Nummerbeteckningar
- LIBRIS-ID:oai:DiVA.org:uu-151579
- ISBN:9789155480783
- https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-151579URI
Kompletterande språkuppgifter
- Språk:engelska
- Sammanfattning på:engelska
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- SwePubSwePub
Klassifikation
Serie
- Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine,1651-6206 ;674
Anmärkningar
- Intra- and intercellular information is communicated by posttranslational modifications (PTMs) and protein-protein interactions, transducing information over cell membranes and to the nucleus. A cells capability to respond to stimuli by several highly complex and dynamic signaling networks provides the basis for rapid responses and is fundamental for the cellular collaborations required in a multicellular organism. Having received diverse stimuli, being positioned at various stages of the cell cycle or, for the case of cancer, containing altered genetic background, each cell in a population is slightly different from its neighbor. However, bulk analyses of interactions will only reveal an average, but not the true variation within a population. Thus studies of interacting endogenous biomolecules in situ are essential to acquire a comprehensive view of cellular functions and communication. In situ proximity ligation assay (in situ PLA) was developed to investigate individual endogenous protein-protein interactions in fixed cells and tissues and was later applied for detection for PTMs. Progression of signals in a pathway can branch out in different directions and induce expression of different target genes. Hence simultaneous measurement of protein activity and gene expression provides a tool to determine the balance and progression of these signaling events. To obtain this in situ PLA was combined with padlock probes, providing an assay that can interrogate both PTMs and mRNA expression at a single cell level. Thereby different nodes of the signaling pathway as well as drug effects on different types of molecules could be investigated simultaneously. In addition to regulation of gene expression, protein-DNA interactions present a mechanism to manage accessibility of the genomic DNA in an inheritable manner, providing the basis for lineage commitment, via e.g. histone PTMs. To enable analyses of protein-DNA interactions in situ we developed a method that utilizes the proximity dependence of PLA and the sequence selectivity of padlock probes. This thesis presents new methods providing researchers with a set of tools to address cellular functions and communication in complex microenvironments, to improve disease diagnostics and to contribute to hopefully finding cures.
Ämnesord och genrebeteckningar
- MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Cell- och molekylärbiologi hsv//swe
- MEDICAL AND HEALTH SCIENCES Basic Medicine Cell and Molecular Biology hsv//eng
- proximity ligation
- in situ PLA
- padlock probe
- rolling circle amplification
- flow cytometry
- in situ
- single cell
- single molecule
- protein interaction
- protein-DNA interaction
- posttranslational modification
- Molecular medicine
- Molekylär medicin
- Molekylär medicin
- Molecular Medicine
Biuppslag (personer, institutioner, konferenser, titlar ...)
- Söderberg, Ola,Associate ProfessorUppsala universitet,Molekylära verktyg (preses)
- Landegren, Ulf,ProfessorUppsala universitet,Molekylära verktyg (preses)
- Larijani, Banafshe,ProfessorCell Biophysics Laboratory Cancer Research UK, Lincolns Inn Fields Laboratories, London Research Institute (opponent)
- Uppsala universitetMolekylära verktyg (creator_code:org_t)
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