Non-Viral Gene Delivery Vectors: Methods and Protocols (Methods in Molecular Biology)

This volume provides readers with a wide collection of the latest and readily reproducible technical protocols available in the field of non-viral gene delivery vectors. The chapters in this book are organized into three major parts: Part I is a section on conventional bolus gene delivery vectors that introduces typical transfection approaches relying on the addition of transfectants to the cell culture medium where the cells are grown in; Part II covers stimuli-responsive bolus transfectants and topics on gene delivery complexes made of smart polymers or stimuli-responsive polymers that change according to the environment they are in and delivered by dripping into cells; Part III discusses examples of substrate-mediated gene delivery-also termed reverse transfection-and the immobilization of a gene delivery vector onto a surface as opposed to more typical bolus delivery from the medium. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and practical, Non-Viral Gene Delivery Vectors: Methods and Protocols is written for experimentalists, and is an essential part of many laboratory bookshelves. This book will help novice and professionals alike succeed in their research in this field. Part I: Conventional Bolus Gene Delivery Vectors 1. Physical Chemical and Biomolecular Methods for the Optimization of Cationic Lipid-Based Lipoplexes In Vitro for the Gene Therapy Applications Santosh K. Misra and Santanu Bhattacharya 2. Cationic Lipid-Based Nucleic Acid Vectors Emile Jubeli, William P. D. Goldring, and Michael D. Pungente 3. Lipoplexes from Non-Viral Cationic Vectors: DOTAP-DOPE Liposomes and Gemini Micelles Sara Falsini and Sandra Ristori 4. Anionic/Zwitterionic Lipid-Based Gene Vectors of pDNA Ana L. Barrn-Berdn, Emilio Aicart, and Elena Junquera 5. Elaboration and Physicochemical Characterization of Niosome-Based Nioplexes for Gene Delivery Purposes Edilberto Ojeda, Mireia Agirre, Ilia Villate-Beitia, Mohamed Mashal, Gustavo Puras, Jon Zarate, and Jose L. Pedraz 6. Quantitative Intracellular Localization of Cationic Lipid-Nucleic Acid Nanoparticles with Fluorescence Microscopy Ramsey N. Majzoub, Kai K. Ewert, and Cyrus R. Safinya 7. Targeted Delivery of Peptide-Tagged DNA Lipoplexes to Hepatocellular Carcinoma Cells Mario Ariatti 8. Lipoplexes Strengthened by Anionic Polymers: Easy Preparation of Highly Effective siRNA Vectors Based on Cationic Lipids and Anionic Polymers Danielle Arruda, Anne Schlegel, Pascal Bigey, and Virginie Escriou 9. Polymer Based Gene Silencing: In Vitro Delivery of SiRNA Margarida I. Simo Carlos, Andreas Schtzlein, and Ijeoma Uchegbu 10. Polyallylamine Derivatives: Novel Non-Toxic Transfection Agents Magdalena Wytrwal and Chantal Pichon 11. Biodegradable Three-Layered Micelles and Injectable Hydrogels Daniel G. Abebe, Rima Kandil, Teresa Kraus, Maha Elsayed, Tomoko Fujiwara, and Olivia M. Merkel 12. Cationic Lipid-Coated Polyplexes ("Lipopolyplexes") for DNA and Small RNA Delivery Alexander Ewe and Achim Aigner 13. Preparation of Targeted Anionic Lipid-Coated Polyplexes for MicroRNA Delivery Xiaomeng Huang, Mengzi Zhang, Xinmei Wang, L. James Lee, and Robert J. Lee Part II: Stimuli-Responsive Bolus Gene Delivery Vectors 14. Characterization and Investigation of Redox-Sensitive Liposomes for Gene Delivery Daniele Pezzoli, Elena Tallarita, Elena Rosini, and Gabriele Candiani 15. From Artificial Amino Acids to Sequence-Defined Targeted Oligoaminoamides Stephan Morys, Ernst Wagner, and Ulrich Lchelt 16. Gene Delivery Method Using Photo-Responsive Poly(-Amino Ester) as Vectors Nan Zheng, Yang Liu, and Jianjun Cheng 17. Thermo-Responsive Polyplex Micelles with PEG Shells and PNIPAM Layer to Protect DNA Cores for Systemic Gene Therapy Junjie Li, Zengshi Zha, and Zhishen Ge Part III: Substrate-Mediated Gene Delivery 18. Application of Polyethylenimine-Grafted Silicon Nanowire Arrays for Gene Transfection Hongwei Wang, Jingjing Pan, Hong Chen, and Lin Yuan