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Manipulation der Expression neuronaler Gerüstproteine mittels RNA-Interferenz

Manipulation der Expression neuronaler Gerüstproteine mittels RNA-Interferenz

          
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About the Book

Bachelorarbeit aus dem Jahr 2013 im Fachbereich Biologie - Genetik / Gentechnologie, Note: 1,7, Humboldt-Universität zu Berlin (Institut für Biologie), Sprache: Deutsch, Abstract: Diese Bachelorarbeit widmet sich der Klonierung und Expression diverser neuer Vektoren, die einerseits auf Basis des pSuper- (und des pLentiLox 3.7-) Plasmids den Gen-Knockdown von PSD-95, einem neuronalen Gerüstprotein, mittels shRNA-induzierter RNA-Interferenz ermöglichen. Andererseits werden auf pCMV-Tag-, pCherry-/pEYFP-N1 und pLentiLox 3.7-Plasmiden basierende Vektoren konstruiert, die die Einbringung diverser und z. T. Knockdown-resistenter PSD-95-Derivate in Säuger-Zellen erlauben. Das langfristige Ziel ist es, die speziellen Eigenschaften dieser einzelnen Vektoren in einem Konstrukt zu kombinieren: Als Vektor lentiviralen Ursprungs soll er die Möglichkeit des schonenden Gentransfers in sensible Nervenzellen in vitro und in vivo durch Infektion bieten. Innerhalb des Neurons kommt es hierdurch zur Stilllegung des "gesunden", endogenen PSD-95 mittels RNA-Interferenz. Währenddessen wird eine vektorkodierte modifizierte, knockdown-resistente Variante des PSD-95 exprimiert, die in der Zelle die Rolle des endogenen Proteins an dessen statt einnimmt. Liegt ein solcher Vektor erst vor, ist es schnell und auf einfache Art und Weise möglich, zusätzliche Mutationen in das knockdown-resistente PSD-95 einzubringen. Das erlaubt den effektiven Scan verschiedener Mutationen, welche zum Beispiel im potentiellen Zusammenhang mit etwaigen Krankheiten stehen könnten. Die Auswirkungen des mutanten Gens auf die Zellphysis können dann mit verschiedensten Methoden, etwa elektrophysiologisch, weiter untersucht werden.


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Product Details
  • ISBN-13: 9783668984387
  • Publisher: Grin Verlag
  • Publisher Imprint: Grin Verlag
  • Height: 210 mm
  • No of Pages: 68
  • Spine Width: 4 mm
  • Width: 148 mm
  • ISBN-10: 3668984387
  • Publisher Date: 18 Jun 2019
  • Binding: Paperback
  • Language: German
  • Returnable: N
  • Weight: 100 gr


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