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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.
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Methods in Molecular Biology TM VOLUME 229 Lentivirus Gene Engineering Protocols Edited by Maurizio Federico From Lentiviruses to Lentivirus Vectors 3 1 From Lentiviruses to Lentivirus Vectors Maurizio Federico 1. Introduction Although a member of the lentivirus group, the equine infectious anemia virus (EIAV) was the first nonplant virus discovered in the first decade of the 20th century (1), lentiviruses were considered as rather mysterious viruses until the isolation of the human immunodeficiency virus type 1 (HIV-1) occurred at the beginning of 1980s. Lentiviruses are enveloped viruses carrying two copies of single-strand positive (i.e., codifying) RNA and are considered the ethiologic agents of acquired immunodeficiency syndromes for a broad range of animal species, such as humans, primates, cats, horses, sheep, and goats. Such syndromes develop in multiorgan diseases and share a long period of incubation (with viral persistence despite a potent immunological response) and a fata