33 - Use of Retroviral Vectors for Gene Transfer and Expression

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Retroviruses have evolved a highly efficient gene transfer capability that provides the basis for one of the most effective gene transfer systems. The retroviral vector system has proved useful for the transfer of genes into many cell types, such as hematopoietic cells and other primary cells that are difficult to transduce by using other methods. In addition, the precise integration of retroviral vectors into the genome of an infected cell provides a powerful genetic tool. Many elegant applications of the retroviral vector system have been developed based on these properties — such as promoter trap vectors for the analysis of cellular promoters and their regulation and the use of retroviral vectors to study immunoglobulin rearrangement. This chapter focuses on the use of high-titer retroviral vectors that are developed for the transfer and expression of complimentary DNAs (cDNAs). The retroviral vector system for gene transfer can be divided into two components, the retroviral vector and the packaging cells. The retroviral vector is manipulated in its DNA form as part of a bacterial plasmid. The vector does not encode viral proteins but serves as a vehicle for the genes to be transferred. The retro virus-packaging cells provide all of the viral proteins necessary for encapsidation of vector RNA into virions and for subsequent infection, reverse transcription, and integration of the vector into the genomic DNA of cells.