Lentivirus Gene Engineering Protocols

Maurizio Federico has assembled a panel of outstanding experimenters to detail all the theoretical and practical aspects of lentivirus vector-based gene transfer. The authors demonstrate lentivirus vector production with methods for recovering appropriate producer cells, for producing and titrating lentivirus-containing supernatants, and for detecting transduced cells. The applications of lentivirus vector engineering to different cell types include coverage of lymphocytes, dendritic cells, hematopoietic stem and progenitor cells, mesenchymal stem cells, hippocampal neurons, cardiomyocytes, as well as airway epithelia, corneal cells, and retinal pigment. Also advanced are applications in which genetic material is directly inoculated with lentivirus vectors in such experimental animal models as mouse, rat, and rabbit.

Table of Contents:
Part I. Lentivirus Vectors From Lentiviruses to Lentivirus Vectors Maurizio Federico The Choice of a Suitable Lentivirus Vector: Transcriptional Targeting Francesco Lotti and Fulvio Mavilio Choice and Use of Appropriate Packaging Cell Types Annmarie L. Pacchia, Sayandip Mukherjee, and Joseph P. Dougherty Small- to Large-Scale Production of Lentivirus Vectors Michael P. Marino, Milson J. Luce, and Jakob Reiser Detection and Titration of Lentivirus Vector Preparations Joel Gatlin, Miguel Islas-Ohlmayer, and J. Victor Garcia Detection and Selection of Lentiviral Vector-Transduced Cells Yan Cui and Lung-Ji Chang Detection of Replication-Competent Lentiviral Particles Harry Segall and Richard E. Sutton Part II. Transduction of Ex Vivo Cell Cultures Through Lentivirus Vectors Lymphocytes Dhanalakashmi Chinnasamy and Fabio Candotti Monocyte/Macrophages and Dendritic Cells Harry Segall and Richard E. Sutton Hematopoietic Stem and Progenitor Cells Robert E. Donahue and Irvin S. Y. Chen Mesenchymal Stem Cells Xian-Yang Zhang, Vincent F. La Russa, and Jakob Reiser Hippocampal Neurons Jay L. Nadeau Dorsal Root Ganglia Sensory Neurons Jane Fleming, Samantha L. Ginn, and Ian E. Alexander Cardiomyocytes Desiree Bonci, Michael V. G. Latronico, and Gianluigi Condorelli Airway Epithelia Larry G. Johnson, Scott H. Randell, and John C. Olsen Corneal Cells Binoy Appukuttan, Trevor McFarland, and J. Timothy Stout Retinal Tissue Trevor McFarland, Binoy Appukuttan, and J. Timothy Stout Part III. Advantages and Uses of Non-HIV-1-Based Lentivirus Vectors HIV-2 Vectors Suresh K. Arya, Jean E. Cho, and Suzie H. Chang SIV Vectors Alessia Ruggieri, Didier Negre, and Francois-Loic Cosset FIV Vectors Nils Loewen, Roman Barraza, Todd Whitwam, Dyana T. Saenz, Iris Kemler, and Eric M. Poeschla Hybrid Lentivirus Vectors, Vicente Planelles Part IV. Direct In Vivo Transduction by Lentivirus Vector and Detection Cells of Respiratory Epithelium Patrick L. Sinn and Paul B. McCray, Jr. Cells of the Nervous System Carl Rosenblad and Cecilia Lundberg Index