UNCORRECTED PROOF CHAPTER EIGHTEEN C0090 Genetic Nanomedicine: Gene Delivery by Targeted Lipoplexes Nejat Du ¨zgu ¨nes ¸* and Conchita Tros de Ilarduya † Contents 1. Introduction 2 2. Preparation of Protein–Cationic Lipid–DNA Ternary Complexes 3 2.1. Liposome preparation 3 2.2. Plasmids 4 2.3. Lipoplex preparation 4 3. Gene Delivery In Vitro 5 4. Cell Viability Following Transfection 7 5. Enhancement of Transfection In Vitro 8 6. Targeted Lipoplexes In Vivo 10 7. Concluding Remarks 11 Acknowledgments 11 References 12 Abstract sp0025 Cationic liposome–DNA complexes (lipoplexes) are used for the delivery of plasmid DNA to cultured cells and various tissues in vivo. In this chapter, we describe the preparation and evaluation of plain and targeted lipoplexes, using targeting ligands, including epidermal growth factor and transferrin. Ligand- associated lipoplexes may be used to target DNA or other nucleic acid drugs to specific cells, particularly cancer cells that overexpress the receptors for the ligands. We provide examples of the enhancement of gene expression mediated by epidermal growth factor in murine, human oral squamous cell carcinoma cells, human hepatoblastoma, and rat colon adenocarcinoma cells. We also summarize the studies on the use of transferrin–lipoplexes for enhancing gene delivery to cervical carcinoma, murine colon carcinoma, and African green monkey kidney cells. We outline two animal models in which transferrin– Comp. by: GKarthikeyan Stage: Proof Chapter No.: 18 Title Name: MIE Date:6/3/12 Time:12:33:41 Page Number: 1 Methods in Enzymology, Volume 509 # 2012 Elsevier Inc. ISSN 0076-6879, DOI: 10.1016/B978-0-12-391858-1.00018-6 All rights reserved. * Department of Biomedical Sciences, Arthur A. Dugoni School of Dentistry, University of the Pacific, San Francisco, California, USA { Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, University of Navarra, Pamplona, Spain 1 MIE, 978-0-12-391858-1 B978-0-12-391858-1.00018-6, 00018 To protect the rights of the author(s) and publisher we inform you that this PDF is an uncorrected proof for internal business use only by the author(s), editor(s), reviewer(s), Elsevier and typesetter SPi. It is not allowed to publish this proof online or in print. This proof copy is the copyright property of the publisher and is confidential until formal publication.