Contents -- Preface -- Contributors -- 1. Non-Viral Gene Therapy Sean M. Sullivan -- 1. Introduction -- 2. Plasmid DNA -- 2.1 Plasmid DNA Manufacture -- 3. Plasmid DNA Gene Transfer Methods -- 3.1 Plasmid DNA or "Naked DNA" as a Gene Delivery System -- 3.1.1 Electroporation of Naked DNA -- 3.1.2 Sonoporation of Naked DNA -- 3.2 Plasmid DNA Formulations -- 3.2.1 Cationic Lipids -- 3.2.1.1 In vitro transfection -- 3.2.1.2 Systemic in vivo gene transfer -- 3.2.1.3 Local administration of cationic lipid/pDNA transfection complexes -- 3.3 Polymer -- 3.3.1 Cationic Polymers -- 3.3.2 Neutral Polymer -- Conclusions -- References -- 2. Adenoviral Vectors Stuart A. Nicklin and Andrew H. Baker -- 1. Introduction -- 2. Adenoviral Capsid Structure -- 3. Adenoviral Cell Entry -- 4. Production of Adenoviral Vectors -- 5. Production of Targeted Adenoviral Vectors -- 6. Gene Therapy Applications -- 7. Immune Responses to Ad Vectors -- 8. Safety and Regulatory Issues -- 9. Conclusions -- References -- 3. Retroviral Vectors and Integration Analysis Cynthia C. Bartholomae, Romy Kirsten, Hanno Glimm, Manfred Schmidt and Christof von Kalle -- 1. Introduction -- 2. Design, Production and Mechanism of Transduction -- 3. In vivo Application -- 4. Side Effects in Retroviral Gene Therapy -- 4.1 Distribution of Retroviral Integration Sites in the Cellular Genome -- 4.2 Side Effects in Clinical and Preclinical Gene Therapy Studies -- 5. New Strategies for Vector Biosafety in Gene Therapy -- References -- 4. Lentiviral Vectors Janka Mátrai, Marinee K. L. Chuah and Thierry VandenDriessche -- 1. Basic Viral Biology -- 2. Vector Design and Production -- 2.1 Vector Development -- 2.2 Vector Production -- 3. Gene Transfer Concepts and Potential Applications -- 3.1 Target Cells and Diseases -- 3.2 Pseudotyping -- 3.3 Cell Type Specific Targeting.

3.4 Integration-Defective Lentiviral Vectors -- 4. Immune Consequences -- 5. Safety Issues -- 6. Conclusions and Perspectives -- References -- 5. Herpes Simplex Virus Vectors William F. Goins, David M. Krisky, James B. Wechuck, Darren Wolfe, Justus B. Cohen and Joseph C. Glorioso -- 1. Introduction -- 2. HSVBiology in the Design of Replication DefectiveVectors -- 3. HSV Vector Design Technology -- 4. Gene Transfer/Therapy Applications -- 5. Immunology -- 6. Safety and Regulatory Issues -- 7. Summary -- References -- 6. Adeno-Associated Viral (AAV) Vectors Nicholas Muzyczka -- 1. Introduction -- 2. Biology of AAV -- 3. Vector Technology -- 4. Vector Characteristics In Vivo -- 5. Next Generation Vectors -- 6. Conclusions and Outlook -- References -- 7. Regulatory RNA in Gene Therapy Alfred. S. Lewin -- 1. Introduction -- 2. Delivery of Therapeutic RNAs -- 3. Ribozymes -- 4. RNAi for Gene Therapy -- 5. Gene Therapy Using miRNA -- 6. Aptamers, Decoys and Bi-Functional RNAs -- 7. Modification of Cis-Acting Regulatory RNA Sequences -- 8. Conclusions -- References -- 8. DNA Integrating Vectors (Transposon, Integrase) Lauren E. Woodard and Michele P. Calos -- 1. Basic Vector Biology -- 1.1 Transposon Systems -- 1.2 Integrase Systems -- 2. Vector Design and Production -- 2.1 Design of Transposon Systems -- 2.2 Design of Integrase Systems -- 2.3 Production of Plasmid DNA -- 3. Gene Transfer Protocols and Potential Applications -- 3.1 Hepatocyte Transfection via Hydrodynamic Injection -- 3.2 Lipophilic Complexes to Transfect Endothelial Cells and Glioblastoma -- 3.3 Direct DNA Injection and Electroporation to Target Muscle, Retina, and Joints -- 3.4 Integration into Cultured Cells for Ex vivo Gene Therapy -- 4. Immunology -- 5. Safety and Regulatory Issues -- 5.1 Integration Profiles and Associated Hazards -- 5.2 Efforts to Enhance Integration Specificity.

5.3 Effects on Tumor Latency in Mouse Models of Cancer -- References -- 9. Homologous Recombination and Targeted Gene Modification for Gene Therapy Matthew Porteus -- 1. Introduction -- 2. Problems with Using Gene Targeting by Homologous Recombination -- 3. Homologous Recombination in Embryonic Stem Cells -- 4. Homologous Recombination using Adeno-Associated Virus -- 5. Site-Specific Modification of the Genome using Double-Strand Breaks -- 6. Double-Strand Break Repair -- 7. Double-Strand Break Induced Homologous Recombination -- 8. Re-design of Homing Endonucleases to Recognize New Target Sites -- 9. Development of Zinc Finger Nucleases -- 10. Using Zinc Finger Nucleases to Stimulate Gene Targeting -- 11. Using Zinc Finger Nucleases to Site-Specifically Modify Genes by Mutagenic Non-Homologous End-Joining -- 12. Strategies of Zinc Finger Nuclease Design -- 13. Aspects of Zinc Finger Binding Sites and Structure of Zinc Finger Nucleases -- 14. Zinc Finger Nuclease Toxicity: Measuring and Minimizing -- 15. The Challenge of Delivery -- 16. Future Directions and Promise of Homologous Recombination as a Gene Correction Approach to Gene Therapy -- References -- 10. Gene Switches for Pre-Clinical Studies in Gene Therapy Caroline Le Guiner, Knut Stieger, Alice Toromanoff, Fabienne Rolling, Philippe Moullier and Oumeya Adjali -- 1. Introduction -- 2. Rapamycin-Dependent Regulatable System -- 2.1 Molecular Mechanisms Involved in Transgene Regulation -- 2.2 Pharmacology of Rapamycin -- 2.3 Translation Development of the Rapamycin Dependent Regulation System . . -- 3. Tetracycline-Dependent Regulatable Systems -- 3.1 Molecular Mechanisms Involved in Transgene Regulation -- 3.2 Pharmacology of Doxycycline (Dox) -- 3.3 Translational Development of Tet-dependant Regulation Systems -- 4. Other Regulatable Systems -- 5. General Conclusions -- References.

11. Gene Therapy for Central Nervous System Disorders Deborah Young and Patricia A. Lawlor -- 1. Introduction -- 2. Gene Therapy for Parkinson's Disease -- 3. Gene Therapy for Temporal Lobe Epilepsy -- 4. Huntington's Disease Gene Therapy -- 5. Amyotrophic Lateral Sclerosis (ALS) -- 6. Gene Therapy for Canavan Disease -- 7. Gene Therapy for Alzheimer's Disease -- 8. Conclusions and Outlook -- References -- 12. Gene Therapy of Hemoglobinopathies Angela E. Rivers and Arun Srivastava -- 1. Introduction -- 2. β-Thalassemia -- 3. Sickle Cell Disease -- 4. Gene Therapy -- 4.1 Oncoretroviral Vector-Mediated Globin Gene Transfer -- 4.2 Lentiviral Vector-Mediated Globin Gene Transfer -- 4.3 Adeno-Associated Viral Vector-Mediated Globin Gene Transfer -- References -- 13. Gene Therapy for Primary Immunodeficiencies Aisha Sauer, Barbara Cassani and Alessandro Aiuti -- 1. Introduction -- 2. Adenosine Deaminase (ADA)-deficient SCID -- 3. X-linked Severe Combined Immunodeficiency (SCID-X1) -- 4. Gene Therapy for Other SCIDs -- 4.1 V(D)J Recombination Defects -- 4.2 Purine Nucleoside Phosphorylase (PNP) Deficiency -- 4.3 Janus Kinase 3 (Jak3) Deficiency -- 4.4 IL-7R Deficiency -- 4.5 Zeta Associated 70 kDa Phosphoprotein (ZAP-70) Deficiency -- 5. Wiskott-Aldrich-Syndrome (WAS) -- 6. Chronic Granulomatous Disease -- 7. Conclusions and Outlook -- References -- 14. Gene Therapy for Hemophilia David Markusic, Babak Moghimi and Roland Herzog -- 1. Introduction -- 2. Limitations of Hemophilia TreatmentWith Coagulation Factor Concentrates or Recombinant Coagulation Factors -- 3. Gene Transfer for Correction of Hemophilia -- 3.1 Ex Vivo Gene Transfer of F.VIII and F.IX -- 3.2 In Vivo Gene Transfer of F.VIII and F.IX . -- 4. AAV is a Preferred Gene Therapy Vector for In Vivo Gene Transfer to Correct of Hemophilia.

5. Immunological Considerations for Efficient F.IX Gene Transfer -- 6. Advancements from Small and Large Animal Models of Hemophilia -- 6.1 Murine Hemophilia Models -- 6.2 Canine Hemophilia Models -- 7. Gene Therapy Trials for Hemophilia Past, Present, and Future -- 8. Conclusions -- References -- 15. Gene Therapy for Obesity and Diabetes Sergei Zolotukhin and Clive H. Wasserfall -- 1. Introduction -- 2. Understanding Obesity: WhyWe Get Fat -- 2.1 Genetic Factors: Human Obesity Gene Map -- 2.2 Environmental Factors: The Big Two and Other Causal Contributors -- 3. General Strategies in Gene Therapy for Obesity -- 4. Gene Delivery Vehicles -- 5. Gene Targets for Obesity -- 5.1 Leptin -- 5.2 Neurocytokines -- 5.3 AMP-Activated Protein Kinase (AMPK) -- 5.4 Adiponectin -- 5.5 Wnt-10b -- 5.6 Obesity Gene Menu à la Carte -- 5.7 Obesity and Diabetes -- References -- 16. Gene Therapy for Duchenne Muscular Dystrophy Takashi Okada and Shin'ichi Takeda -- 1. Introduction -- 1.1 Background of Duchenne Muscular Dystrophy -- 2. Gene-replacement Strategies using Virus Vectors -- 2.1 Choice of Vector -- 2.2 Modification of the Dystrophin Gene and Promoter -- 2.3 Use of Surrogate Genes -- 3. AAV-Mediated Transduction of Animal Models -- 3.1 Vector Production -- 3.2 Animal Models for the Gene Transduction Study -- 3.3 Immunological Issues of rAAV -- 3.4 Intravascular Vector Administration by Limb Perfusion -- 3.5 Global Muscle Therapies -- 4. Safety and Potential Impact of Clinical Trials -- 5. Development of Alternative Strategies -- 5.1 Design of Read-through Drugs -- 5.2 Modification of mRNA Splicing -- 5.3 Ex Vivo Gene Therapy -- 6. Future Perspectives -- 6.1 Pharmacological Intervention -- 6.2 Capsid Modification -- 7. Conclusions and Outlook -- References -- 17. Cancer Gene Therapy Kirsten A.K. Weigel-Van Aken -- 1. Introduction -- 2. Targeting the Tumor Cell.

2.1 DNA Electroporation.