Several years ago, when the discovery of catalytic RNA was recognized in a public manner,many people asked if new ?elds of therapy would soon be available. Although some tentative positive answers were given,nobody would say with certainty that RNA of various kinds was a truly promising means of altering gene expression. In fact,over the past decade,both our knowledge of RNAs with different functions and the utility of RNA in the inhibition or enhancement of gene expression have occurred with great drama. We proceeded in terms of possible therapeutic tools from RNase P and group I introns through "hammerhead" RNA enzymes, antisense technology, and more recently, to RNAi and its derivatives. A useful practical method of RNA delivery in animals will complete the picture. The diversity of RNA and the varied role of it inside cells and in therapy should be a tremendous challenge for young molecular biologists. This volume will make their task easier. Sidney Altman Sterling Professor of Molecular,Cellular & Devel- mental Biology,Nobel Laureate Department of Molecular,Cellular and Developm- tal Biology Yale University V NGTPR 4/23/05 1:00 PM Page VI VI Foreword Delivery of nucleic acids to cells in an animal remains a challenging problem. It is the major obstacle to success of therapeutic approaches using genes and oli- nucleotides,including siRNAs. Solutions found so far by chemists are satisfactory only for transfection of cells in culture.

Inhaltsangabe1 Molecules for Gene Delivery 1-1 Leaf Huang Recent Progress in Nonviral Gene Delivery 1-2 Kenichi Yoshikawa Physical Chemistry of DNA-Carrier Complex 1-3 Kazuo Maruyama Liposome-Mediated Delivery System 1-4 Tomoko Hashimoto Polymeric Gene Carriers 1-5 Kazunori Kataoka Development of Cationic Block Copolymer-Based Gene Delivery System 1-6 Toshinori Sato Chitosan-Mediated Gene Delivery 1-7 Hidetoshi Arima Dendrimers as DNA Carriers 18 Takuro Niidome Use of Synthetic Peptide for Nonviral Gene Delivery 19 Kazuo Sakurai Oligonucleotide Carrier Based on b1,3Glucans 110 Mahito Nakanishi Biological and Chemical Hybrid Vectors 2 Controlled Gene Delivery 21 Hideyoshi Harashima Pharmacokinetics of Gene Delivery in Cells 22 Takeshi Nagasaki Active DNA Release from Complex 23 Masayuki Yokoyama Controlled Release of DNA using ThermoResponsive Polymers 24 Takeshi Nagasaki Active Transport of Exogenous Gene into Nucleus 25 Masayuki Fujii Controlled Intracellular Localization of Oligonucleotides by Chemical Conjugation 26 Osam Mazda Functional Nucleotide Sequences Capable of Promoting Nonviral Genetic Transfer 27 Yoshiyuki Koyama Shielding of cationic charge of the DNA complex to avoid nonspecific interactions for in vivo gene delivery 28 Shigeru Kawakami In Vivo Gene Transfer by Ligand Modified Gene Carriers 29 Ernst Wagner Optimizing Polyplexes into Systemic Viruses for TumorTargeted Gene Therapy 210 Mariko HaradaShiba Gene Transfer and Target Diseases 211 Hironobu Yanagie Clinical Trials Using NonViral Gene Delivery Systems 3 Current Protocoles of Gene Delivery 31 Mitsuru Hashida Evaluation of Size and ZetaPotential of DNA/Carrier Complex 32 Tomoko Ito Observation of DNA / Carrier Complex under Fluorescence Microscopy 33 Taku Satoh Synthesis of Chemically Modified Chitosan and its Gene Transfection Efficiency 34 Hiroyuki Kamiya Newly Designed DNA Fragments for Gene Correcton 35 Tatsuya Okuda Evaluation of Gene Expression in Vivo after Intraveous and Intraportal Administration of Lipoplexes 36 Takuro Niidome Evaluation of DNA Administrated with Gene Carrier in Organs by Southern Blot Hybridization 37 Fuminori Sakurai Evaluation of Immune Response after Administration of Plasmid DNANonviral Vector Complex 38 Kunt R. Madden Gateway RNAi 4 Design of Genes Based on Current RNA Technology 41 Kazunari Taira Design of Intracellulary Active Ribozymes and siRNA 42 Takanori Yokota siRNAbased Inhibition Specific for Mutant Allele in Autosomal Dominant Diseases: SequenceDependent and Independent Discriminations of Mutant and Wildtype Alleles by siRNA 43 Silvere Petit In Vivo RNA Interference: another tool in the box? 44 Hiroaki Kawasaki Suppression of Gene Expression via Chromatin Remodeling and the siRNAInduced Silencing of Transcription 45 Andrew Miller Intracellular delivery of nucleic acids: differences between transfection and siFection reflect differences between DNA and RNA, and between oligodeoxynucleotides and oligonucleotides 46 Junichi Yano In Vivo Antitumour Activity of a New Cationic Liposome siRNA Complex 47 John Rossi Enhancing RNAi with Synthetic RNA Duplexes