Recombinant Sindbis Virus as an Expression System for Cell Biology

Virus-based systems for the expression of heterologous genes are becoming popular tools for cell biological studies. This chapter outlines the general features of expression vectors based on Sindbis virus, an enveloped plus-stranded RNA virus (SIN vectors). The chapter provides specific examples of the use of SIN vectors for studying the function of polytopic membrane proteins—namely, the facilitative glucose transporters and peripheral membrane proteins belonging to the ras superfamily of guanosine triphosphatase (GTPases). SIN vectors offer efficient infection of a wide range of cells and rapid and reliable expression of heterologous proteins. The recombinant SIN vectors offer several advantages: (1) the genome of Sindbis virus is relatively small and can be fully manipulated by standard in vitro DNA/RNA technology, (2) complete recombinant infectious genomic RNA can be easily constructed in vitro and transfected into host cells to raise viral stocks without selection and plaque purification of recombinant virus, and (3) a variety of SIN vectors and temperature-sensitive mutants can be used to control the level of heterologous gene expression