A novel method to interfere with gene expression in mice using lentiviral transgenesis [Elektronische Ressource] / presented by Milen Kirilov

DissertationSubmitted to theCombined Faculties for the Natural Sciences and for MathematicsOf the Ruperto-Carola University of Heidelberg, GermanyFor the degree ofDoctor of Natural SciencesPresented byMilen KirilovRepublic of BulgariaOral-examinationThemaA NOVEL METHOD TO INTERFERE WITH GENE EXPRESSION IN MICEUSING LENTIVIRAL TRANSGENESIS Referees: Prof. Dr. Günther Schütz Prof. Dr. Renato ParoTABLE OF CONTENTS1. Summary 11. Zusammenfassung 22. Introduction 3 2.1. Nuclear receptor transcription factors 4 2.1.1. Structure of nuclear receptors 4 2.1.2. HNF4γ 5 2.2. RNA interference 8 2.2.1. siRNA and microRNAs (miRNA) 92.2.2. Selection of highly effective siRNA sequences 102.2.3. RISC function 112.2.4. Other silencing models 122.3. Lentiviral transgenesis 132.3.1. Lentiviruses 142.3.2. Lentiviral gene transfer vectors 152.3.3. Self-inactivating vectors 152.3.4. Vector elements 162.3.5. The lentiviral packaging system 162.3.6. Generation of transgenic animals by using lentiviruses 162.3.7. Induction of RNAi in transgenic mice generated by the use of lentiviral vectors 192.3.8. Aim of the project 203. Results 21 3.1. Analysis of HNF4γ expression 213.2.