An inducible RNAi system for the functional dissection of genes in the mouse [Elektronische Ressource] / by Joana Alves Vidigal

An Inducible RNAi system for the functional dissection of genes in the mouse Inaugural-Dissertation to obtain the academic degree Doctor rerum naturalium (Dr. rer. nat.) submitted to the Department of Biology, Chemistry and Pharmacy of the Freie Universität Berlin by Joana Alves Vidigal from Lisbon, Portugal December 2010 The work here described was performed at the Max-Planck Institute for Molecular Genetics in Berlin between February 2007 and December 2010 under the supervision of Prof. Dr. Bernhard G. Herrmann and Dr. Markus Morkel st1 Reviewer: Prof. Dr. Bernhard G. Herrmann Max-Planck Institute for Molecular Genetics Ihnestrasse 73 D-14195 Berlin nd2 Reviewer: Prof. Dr. Stephan Sigrist Institute of Biology - Freie Universität Berlin Takustr. 6, 14195 Berlin Date of defence: 11/02/2011 Aos meus pais e aos meus irmãos Table of Contents 1  INTRODUCTION 11 1.1  An introduction to the mouse as a model system 12 1.2  The molecular revolution of mouse genetics 13 1.3  The “knockout mouse” 14 1.3.1  Isolation of Embryonic Stem Cells 14 1.3.2  Genetic engineering by homologous recombination. 15 1.4  Further developments in transgenic techniques 18 1.4.1  Site-specific recombination systems 18 1.4.2  Inducible expression systems 19 1.4.3  RNAi: an alternative to traditional gene targeting 22 1.5  RNA interference and its triggers 24 1.