The purpose of this volume of Methods in Molecular Medicine is to set forth examples of the great variety of techniques and applications that are now emerging in the field of nonviral gene therapy. The book emphasizes not only specific approaches to gene delivery but, in particular, the best current me- ods to prepare, handle, and characterize gene delivery agents. These topics are of very broad importance since gene therapy evolves from its mostly ac- emy-based experimental and clinical research to the ever increasing number of industry-driven programs directed toward commercial development. S- cessful introduction of nonviral gene therapy agents into the clinic should be expected to require rigorous manufacturing and analytical methods that readily meet the regulatory guidelines under which new drug candidates are reviewed for marketing approval. Exactly what those guidelines will prove to be c- tainly depends on the established guidelines for review of both biological and chemical therapeutics. Additionally, many new techniques are being devised and applied to gene therapy research; these techniques will be instrumental in developing and characterizing successful gene delivery agents. Nonviral Vectors for Gene Therapy: Methods and Protocols has two main sections. To start with, there is a series of chapters on specific protocols for the synthesis, characterization, and application of gene delivery agents. S- eral chapters address the topic of materials to bind with DNA to form the compact condensed phases that facilitate cellular delivery.

Includes supplementary material: sn.pub/extras

Klappentext
Nonviral gene therapy-an emerging field with great clinical promise-avoids many problems associated with viral gene therapy and offers the possibility of superior clinical effectiveness. In Nonviral Vectors for Gene Therapy: Methods and Protocols, Mark A. Findeis has assembled a panel of active researchers to present their best methods not only for preparing, handling, and characterizing gene delivery agents, but also for gene delivery. To help those preparing and characterizing gene transfer agents, the contributors examine a broad range of compounds that bind with DNA to form the compact complexes that facilitate cellular delivery-among them peptide conjugates, synthetic polymers, and lipids. They also outline specific approaches to gene transfer in vivo, including direct delivery by intratumoral injection and indirect delivery by cell-specific targeting of DNA complexes, and discuss in detail many spectroscopic techniques for characterizing nonviral gene delivery agents. Opportunities for the development and application of these novel vectors in the research lab and eventually, in the clinic, are highlighted.
Comprehensive and state-of-the-art, Nonviral Vectors for Gene Therapy: Methods and Protocols illuminates for today's investigators the powerful new approaches to the creation of nonviral materials, as well as their efficacious use in delivering therapeutic genes to patients across the spectrum of human disease.

Inhalt
Synthesis of Polyampholyte Comb-Type Copolymers Consisting of a Poly(L-lysine) Backbone and Hyaluronic Acid Side Chains for DNA Carrier.- Cationic ?-Helical Peptides for Gene Delivery into Cells.- Supramolecular Self-Assembly of Poly(ethylene glycol)-Block-Poly (L-lysine) Dendrimer with Plasmid DNA.- Water-Soluble Cationic Methacrylate Polymers for Nonviral Gene Delivery.- Stabilization of Polycation-DNA Complexes by Surface Modification with Hydrophilic Polymers.- Use of Disulfide Cationic Lipids in Plasmid DNA Delivery.- Interactions of Lipid-Oligonucleotide Conjugates with Low-Density Lipoprotein.- Coupling of Nuclear Localization Signals to Plasmid DNA.- Progress Toward a Synthetic Virus.- Characterization of Polyampholyte Comb-Type Copolymer DNA Carriers.- Methods for Studying the Formation of Polycation-DNA Complexes and Properties Useful for Gene Delivery.- Characterization of DNA Condensates by Atomic Force Microscopy.- Rapid and Systematic Transfer and Recovery of Large BACs/PACs into Mammalian Cells by HAEC Retrofitting.- Systemic Delivery of Therapeutic Proteins by Intramuscular Injection of Plasmid DNA.- Local Delivery of Therapeutic Proteins by Intratumoral Injection of Plasmid DNA-Lipid Complexes.- Nonviral DNA Delivery from Polymeric Systems.- Promotion of Duplex and Triplex DNA Formation by Polycation Comb-Type Copolymers.- Lyophilization of Nonviral Gene Delivery Systems.- Ultraviolet Absorption and Circular Dichroism Spectroscopy of Nonviral Gene Delivery Complexes.- Characterization of Synthetic Gene Delivery Vectors by Infrared Spectroscopy.- Characterization of Cationic Vector-Based Gene Delivery Vehicles Using Isothermal Titration and Differential Scanning Calorimetry.- Light-Scattering Techniques for Characterization of Synthetic Gene TherapyVectors.- Renal Gene Therapy.